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2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
2 ATP + 2 cob(II)yrinic acid a,c-diamide + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)yrinic acid a,c-diamide + an oxidized flavoprotein
2 ATP + 2 [corrinoid adenosyltransferase]-cob(I)alamin = 2 triphosphate + 2 adenosylcob(III)alamin + 2 [corrinoid adenosyltransferase]
(1c)
-
-
-
2 ATP + 2 [corrinoid adenosyltransferase]-cob(I)yrinic acid a,c-diamide = 2 triphosphate + 2 adenosylcob(III)yrinic acid a,c-diamide + 2 [corrinoid adenosyltransferase]
(2c)
-
-
-
2 cob(II)alamin + 2 [corrinoid adenosyltransferase] = 2 [corrinoid adenosyltransferase]-cob(II)alamin
(1a)
-
-
-
2 cob(II)yrinic acid a,c-diamide + 2 [corrinoid adenosyltransferase] = 2 [corrinoid adenosyltransferase]-cob(II)yrinic acid a,c-diamide
(2a)
-
-
-
a reduced flavoprotein + 2 [corrinoid adenosyltransferase]-cob(II)alamin = an oxidized flavoprotein + 2 [corrinoid adenosyltransferase]-cob(I)alamin
(1b), spontaneous
-
-
-
a reduced flavoprotein + 2 [corrinoid adenosyltransferase]-cob(II)yrinic acid a,c-diamide = an oxidized flavoprotein + 2 [corrinoid adenosyltransferase]-cob(I)yrinic acid a,c-diamide
(2b), spontaneous
-
-
-
2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
reaction mechanism
-
2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
reaction mechanism, substrate structure
-
2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
active site and substrate binding sites and structures, molecular architecture
-
2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
Cys79, Cys80, and Cys83 are important for catalysis
-
2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
nucleophilic attack from reduced Co1+ ion of cob(I)alamin to the C-5 carbon of ATP, ordered substrate binding mechanism with ATP being first and essential for catalysis
-
2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
(1) overall reaction
-
-
-
2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
residues Phe91 and Trp93 play a critical role in the mechanism of four-coordinate Cob(II)alamin formation in the active site of the Salmonella enterica ATP:Co(I)rrinoid adenosyltransferase enzyme, overview. The enzyme adopts a closed conformation and residues Phe91 and Trp93 displace 5,6-dimethylbenzimidazole, the lower nucleotide ligand base of cobalamin, to generate a transient four-coordinate cobalamin, which is critical in the formation of the AdoCbl Co-C bond, important role of bulky hydrophobic side chains in the active site. CobA and PduO increase the redox potential of the cob(II)alamin/cob(I)alamin couple to facilitate formation of the Co-C bond, in both cases the polar coordination of the lower ligand to the cobalt ion is eliminated by placing that face of the corrin ring adjacent to a cluster of bulky hydrophobic side chains
-
2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
to overcome the thermodynamically challenging Co2+ -> Co1+ reduction, the enzyme drastically weakens the axial ligand-Co2+ bond so as to generate effectively four-coordinate Co2+-corrinoid species, mechanism, overview. The entire hydrophobic pocket below the corrin ring, and not just residue F112, is critical for the removal of the axial ligand from the cobalt center of the Co2+-corrinoids. Large role of the ATP-induced active-site conformational changes with respect to the formation of 4c Co(II)Cbl
-
2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
active site and substrate binding sites and structures, molecular architecture
-
-
2 ATP + 2 cob(II)yrinic acid a,c-diamide + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)yrinic acid a,c-diamide + an oxidized flavoprotein
reaction mechanism
-
2 ATP + 2 cob(II)yrinic acid a,c-diamide + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)yrinic acid a,c-diamide + an oxidized flavoprotein
active site and substrate binding sites and structures, molecular architecture
-
2 ATP + 2 cob(II)yrinic acid a,c-diamide + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)yrinic acid a,c-diamide + an oxidized flavoprotein
Cys79, Cys80, and Cys83 are important for catalysis
-
2 ATP + 2 cob(II)yrinic acid a,c-diamide + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)yrinic acid a,c-diamide + an oxidized flavoprotein
not yet confimed
-
2 ATP + 2 cob(II)yrinic acid a,c-diamide + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)yrinic acid a,c-diamide + an oxidized flavoprotein
not yet confimed
2 ATP + 2 cob(II)yrinic acid a,c-diamide + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)yrinic acid a,c-diamide + an oxidized flavoprotein
residues Phe91 and Trp93 play a critical role in the mechanism of four-coordinate Cob(II)alamin formation in the active site of the Salmonella enterica ATP:Co(I)rrinoid adenosyltransferase enzyme, overview.. The enzyme adopts a closed conformation and residues Phe91 and Trp93 displace 5,6-dimethylbenzimidazole, the lower nucleotide ligand base of cobalamin, to generate a transient four-coordinate cobalamin, which is critical in the formation of the AdoCbl Co-C bond, important role of bulky hydrophobic side chains in the active site. CobA and PduO increase the redox potential of the cob(II)alamin/cob(I)alamin couple to facilitate formation of the Co-C bond, in both cases the polar coordination of the lower ligand to the cobalt ion is eliminated by placing that face of the corrin ring adjacent to a cluster of bulky hydrophobic side chains
-
2 ATP + 2 cob(II)yrinic acid a,c-diamide + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)yrinic acid a,c-diamide + an oxidized flavoprotein
(2) overall reaction
-
-
-
2 ATP + 2 cob(II)yrinic acid a,c-diamide + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)yrinic acid a,c-diamide + an oxidized flavoprotein
active site and substrate binding sites and structures, molecular architecture
-
-
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2 ATP + 2 cob(II)alamin + a reduced flavoprotein
2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
2'-deoxy-ATP + cob(I)alamin + H2O
?
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(I)alamine
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
ATP + cob(I)alamin
triphosphate + adenosylcobalamin
ATP + cob(I)alamin
triphosphate + coenzyme B12
ATP + cob(I)alamin
tripolyphosphate + coenzyme B12
ATP + cob(I)alamin + H2O
?
anaerobic, 20 min, 80°C, pH 8, in presence of 1 mM titanium (III) citrate
ATP-dependent cob(I)alamin consumption to a yet unknown compound
-
?
ATP + cob(I)alamin + H2O
adenosylcobalamin + diphosphate + phosphate
-
37°C
monitoring adenosylcobalamin formation at 388 nm in continous spectrophotometric assay
-
?
ATP + cob(I)alamin + H2O
phosphate + diphosphate + adenosylcobalamin
anaerobic, 20 min, 80°C, pH 8, in presence of 1 mM titanium (III) citrate
measured by decrease in light absorbance by cob(I)alamin at 388 nm and increase of light absorbance by presumably adenosylcobalamin at 525 nm
-
?
ATP + cob(I)alamine
triphosphate + adenosylcob(I)alamine
-
-
-
?
ATP + cob(I)yrinic acid a,c-diamide
triphosphate + adenosylcob(III)yrinic acid a,c-diamide
-
-
-
?
ATP + cob(II)alamin
triphosphate + adenosylcob(II)alamine
ATP + cob(II)alamine
triphosphate + adenosylcob(II)alamine
-
-
-
?
ATP + cob(II)inamide
triphosphate + adenosylcob(II)inamide
-
-
-
?
ATP + cobalamin
triphosphate + adenosylcob(III)alamin
-
enzyme is absolutely specific for ATP or dATP as adenosyl donors, ATP is the preferred adenosyl donor
-
-
?
ATP + cobalamin
triphosphate + adenosylcobalamin
enzyme is absolutely specific for ATP or dATP as adenosyl donors
-
-
ir
ATP + cobinamide
triphosphate + adenosylcobinamide
ATP + hydroxocobalamin
?
-
-
-
?
ATP + hydroxycobalamin
adenosylcobalamin + phosphate + diphosphate
37°C, pH 8, 0.5 mM ATP, 0.05 mM hydroxycobalamin, in presence of 1 mM titanium(III)citrate
measured by decrease in absorbance at 388 nm
-
?
ATP + hydroxycobalamin
tripolyphosphate + adenosylcobalamin + H2O
-
coenzyme B12 synthesis from vitamin B12, dimethylbenzimidazole arm of vitamin B12 plays no role in substrate positioning, corrinoid adenosylation assay: anaerobic, pH 6, 25°C, 1 or 2 mM FMN, 10 or 20 mM NADH, NAD(P)H: flavin oxidoreductase, 2 h incubation for complete reduction of hydroxycobalamin to cob(II)alamin before initiation of adenosyltransfer
measuring difference in absorbance by adenosylcobalamin at 525 nm
-
?
cob(I)alamin + ADP
adenosylcobalamin + diphosphate
cob(I)alamin + ATP
adenosylcobalamin + ?
-
in the co+ assay the cobalt ion of cobalamin is chemically reduced in solution to cob(I)alamin by Ti(III)citrate, allowing the cob(I)alamin adenosylation reaction to be measured directly. The Co+ assay is performed under anoxic conditions
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
cob(I)alamin + ATP + H2O + H+
adenosylcobalamin + diphosphate + phosphate
cob(I)alamin + CTP + H2O + H+
cytosylcobalamin + diphosphate + phosphate
-
-
-
-
?
cob(I)alamin + dATP + H2O + H+
deoxyadenosylcobalamin + diphosphate + phosphate
-
-
-
-
?
cob(I)alamin + GTP + H2O + H+
guanosylcobalamin + diphosphate + phosphate
-
low activity
-
-
?
cob(I)alamin + ITP
hypoxanthosylcobalamin + diphosphate + phosphate
-
low activity
-
-
?
cob(I)alamin + UTP
uranylcobalamin + diphosphate + phosphate
-
low activity
-
-
?
cob(I)inamide + ATP
5'-deoxy-5'-adenosyl-cob(I)inamide + polyphosphate
-
-
?
cob(I)inamide + ATP
adenosylcobinamide + triphosphate
cob(I)yric acid + ATP
5'-deoxy-5'-adenosyl-cob(I)yric acid + polyphosphate
-
-
?
cob(I)yrinic acid a,c-diamide + ATP
5'-deoxy-5'-adenosyl-cob(I)yrinic acid a,c-diamide + polyphosphate
-
-
?
cob(II)alamin
cob(I)alamin
G97, T161, and H183 possible role in stabilizing four-coordinate, cob(II)alamin C-terminal His-tagged enzyme binds cob(II)alamin base-off while N-terminal His-tagged enzyme binds it base-on (impaired base-off transition), only mutants S68F, K78Q, K78R, R186W, and R190C also bind cob(II)alamin base-off
-
-
?
cob(II)alamin + ATP
adenosylcobalamin + ?
-
in the Co2+ assay the NADPH-dependent flavodoxin protein reductase/flavodoxin system is used to reduce Co2+ to Co+. In this Co2+ assay, the PduO enzyme must bind cob(II)alamin and facilitate the generation of cob(I)alamin in its active site
-
-
?
CTP + cob(I)alamin
triphosphate + cytosylcobalamin
-
polymorphic variants 239K and 239M, 9% activity compared to ATP with enzyme variant 239K, 6% activity compared to ATP with enzyme variant 239M
-
-
?
CTP + cob(I)alamin + H2O
cytidylcobalamin + diphosphate + phosphate
-
37°C
-
-
?
cyanocob(I)alamin + ATP
tripolyphosphate + alpha-(5,6-dimethylbenzimidazolyl)deoxyadenosylcobamide
dATP + cob(I)alamin
tripolyphosphate + deoxyadenosylcobalamin
-
-
?
dATP + cobalamin
triphosphate + deoxyadenosylcob(III)alamin
-
enzyme is absolutely specific for ATP or dATP as adenosyl donors, dATP results in 21% of the activity with ATP
-
-
?
dATP + cobalamin
triphosphate + deoxyadenosylcobalamin
enzyme is absolutely specific for ATP or dATP as adenosyl donors
-
-
ir
GTP + cob(I)alamin
triphosphate + guanosylcobalamin
-
polymorphic variants 239K and 239M, 16% activity compared to ATP with enzyme variant 239K, 14% activity compared to ATP with enzyme variant 239M
-
-
?
GTP + cob(I)alamin + H2O
guanosylcobalamin + diphosphate + phosphate
-
37°C
-
-
?
ITP + cob(I)alamin + H2O
inosylcobalamin + diphosphate + phosphate
-
37°C
-
-
?
UTP + cob(I)alamin
triphosphate + uridylcobalamin
-
polymorphic variants 239K and 239M, 8% activity compared to ATP with enzyme variant 239K, 6% activity compared to ATP with enzyme variant 239M
-
-
?
UTP + cob(I)alamin + H2O
uridylcobalamin + diphosphate + phosphate
-
37°C
-
-
?
additional information
?
-
2 ATP + 2 cob(II)alamin + a reduced flavoprotein
2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
-
-
-
?
2 ATP + 2 cob(II)alamin + a reduced flavoprotein
2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
-
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(I)alamine
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(I)alamine
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(I)alamine
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
-
adenosyltransferase enzymes lower the thermodynamic barrier of the Co2+ toCo+ reduction needed for the formation of the unique organometalic Co-C bond of adenosylcobalamin
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
reaction mechanism: assimilated cobalamin is reduced to co(II)alamin, that then binds to the enzyme-ATP complex, further reduction yields a nucleophilic four coordinated Co1+ intermediate that attacks the 5'-carbon of the cosubstrate ATP to generate adenosylcobalamin and triphosphate, spectroscopic analysis, enzyme-induced base-on/base-off conversion activating the cobalamin substrate for reduction
-
-
ir
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
-
adenosyltransferase enzymes lower the thermodynamic barrier of the Co2+ to Co+ reduction needed for the formation of the unique organometalic Co-C bond of adenosylcobalamin
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
binding of the substrate ATP to ATR that is fully loaded with 5'-deoxyadenosylcobalamin leads to the ejection of 1 equivalent of the cofactor into solution. In the presence of methylmalonyl-CoA mutase and ATP, 5'-deoxyadenosylcobalamin is transferred from ATR to the acceptor protein in a process that exhibits an 3.5fold lower Kact for ATP compared to the one in which cofactor is released into solution. ATP favorably influences cofactor transfer in the forward direction by reducing the ratio of apo-methylmalonyl-CoA mutase/holo-ATR required for delivery of 1 equivalent of 5'-deoxyadenosylcobalamin, from 4 to 1. A rotary rotary mechanism for ATR function is proposed in which, at any given time, only two of its active sites are used for 5'-deoxyadenosylcobalamin synthesis and where binding of ATP to the vacant site leads to the transfer of the high value 5'-deoxyadenosylcobalamin product to the acceptor mutase
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
final step in the conversion of vitamin B12 to coenzyme B12, the latter being required for degradation of 1,2-propanediol
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
last step in the corrinoid adenosylation pathway, EutT and CobA
-
-
ir
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
mechanism of adenosylcobalamin biosynthesis
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
CobA has an ATP-binding P-loop motif, while EutT has a cysteine-rich region reminiscent of a conserved S-adenosylmethionine Fe-S cluster motif
-
-
ir
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
formation of the the essential C-Co bond by transferring the adenosyl group from a molecule of ATP to a transient Co1+ corrinoid species generated by the active site
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
last step in the corrinoid adenosylation pathway, EutT and CobA
-
-
ir
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
CobA has an ATP-binding P-loop motif, while EutT has a cysteine-rich region reminiscent of a conserved S-adenosylmethionine Fe-S cluster motif
-
-
ir
ATP + cob(I)alamin
triphosphate + adenosylcobalamin
-
polymorphic variants 239K and 239M, biosynthesis of adenosylcobalamin
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcobalamin
-
polymorphic variants 239K and 239M, ATP is highly preferred as adenosyl donor
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcobalamin
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
r
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
stereospecific process which proceeds with overall inversion of configuration at C-5' of the adenosyl moiety
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
transfers the adenosyl-group of ATP to the reduced cobalt atom of the cobalamin molecule
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
involved in vitamin B12 metabolism
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
adenosylcobalamin, B12-coenzyme or deoxyadenosyl-B12
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
the main role of this enzyme is apparently the conversion of inactive cobalamins to adenosyl cobalamin for 1,2 propanediol degradation
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
-
?
ATP + cob(I)alamin
tripolyphosphate + coenzyme B12
-
-
-
?
ATP + cob(I)alamin
tripolyphosphate + coenzyme B12
-
-
-
?
ATP + cob(II)alamin
triphosphate + adenosylcob(II)alamine
-
-
-
-
?
ATP + cob(II)alamin
triphosphate + adenosylcob(II)alamine
-
-
-
-
?
ATP + cobinamide
triphosphate + adenosylcobinamide
-
-
-
-
?
ATP + cobinamide
triphosphate + adenosylcobinamide
-
-
-
?
cob(I)alamin + ADP
adenosylcobalamin + diphosphate
-
-
-
-
?
cob(I)alamin + ADP
adenosylcobalamin + diphosphate
-
-
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
-
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
adenosylcobalamin is a cofactor required by the methylmalonyl-CoA mutase
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
cobalamin assimilation and recycling pathway, overview, enzyme deficiency causes methylmalonic aciduria, MMA, is an autosomal recessive disease with symptoms that include ketoacidosis, lethargy, recurrent vomiting, dehydration, respiratory distress, muscular hypotonia and death due to methylmalonic acid levels that are up to 1000fold greater than normal, overview
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
dissociation constant (Kd) of wild-type MMAB for hydroxomethylcobalamin is 0.051 mM and for ATP is 0.365 mM, cobalamin enhances the affinity of MMAB for ATP, while ATP does not show detectable effects on cobalamin binding
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
adenosylation of the corrinoid ring of cob(I)alamin generates coenzyme B12, i.e. adenosylcobalamin or AdoCbl, an essential cofactor used by enzymes that catalyze intramolecular rearrangements, deaminations, dehydrations, reductions, and reductive dehalogenations
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
adenosylation of the corrinoid ring of cob(I)alamin, active site structure, ATP binding motif at the protein N terminus
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
upon binding to LrPduO that is preincubated with ATP, both Co2+corrinoids undergo a partial (40-50%) conversion to distinct paramagnetic Co2+ species. The spectroscopic signatures of these species are consistent with essentially four-coordinate, square-planar Co2+ complexes. For effecting Co2+ to Co1+ reduction the formation of an activated Co2+ corrinoid intermediate that lacks any significant axial bonding interactions is involved to stabilize the redoxactive, Co 3dz2-based molecular orbital
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
adenosylation of the corrinoid ring of cob(I)alamin generates coenzyme B12, i.e. adenosylcobalamin or AdoCbl, an essential cofactor used by enzymes that catalyze intramolecular rearrangements, deaminations, dehydrations, reductions, and reductive dehalogenations
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
adenosylation of the corrinoid ring of cob(I)alamin, active site structure, ATP binding motif at the protein N terminus
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
-
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
cobalamin is a better substrate than cobinamide, the beta-phosphate of ATP is required for binding to the enzyme
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
the enzyme is required to adenosylate de novo biosynthetic intermediates of adenosylcobalamin and to salvage incomplete and complete corrinoids from the environment of this bacterium
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
in vitro reduced flavodoxin provides an electron to generate the co(I)rrinoid substrate in the CobA active site, modeling of enzyme ligand interaction, residues R9 and R165 are important for CobA-FldA docking but not to catalysis, overview
-
-
?
cob(I)alamin + ATP + H2O + H+
adenosylcobalamin + diphosphate + phosphate
-
-
-
-
?
cob(I)alamin + ATP + H2O + H+
adenosylcobalamin + diphosphate + phosphate
-
FMN and NADH are used to reduce cob(III)alamin to cob(I)alamin, the enzyme shows ATP hydrolyzing activity to adenosine and triphosphate in absence of cob(I)alamin
-
-
?
cob(I)inamide + ATP
adenosylcobinamide + triphosphate
-
upon binding to LrPduO that is preincubated with ATP, both Co2+corrinoids undergo a partial (40-50%) conversion to distinct paramagnetic Co2+ species. The spectroscopic signatures of these species are consistent with essentially four-coordinate, square-planar Co2+ complexes. For effecting Co2+ to Co1+ reduction the formation of an activated Co2+ corrinoid intermediate that lacks any significant axial bonding interactions is involved to stabilize the redoxactive, Co 3dz2-based molecular orbital
-
-
?
cob(I)inamide + ATP
adenosylcobinamide + triphosphate
-
-
-
-
?
cob(I)inamide + ATP
adenosylcobinamide + triphosphate
-
cobalamin is a better substrate than cobinamide, the beta-phosphate of ATP is required for binding to the enzyme
-
-
?
cyanocob(I)alamin + ATP
tripolyphosphate + alpha-(5,6-dimethylbenzimidazolyl)deoxyadenosylcobamide
-
-
-
?
cyanocob(I)alamin + ATP
tripolyphosphate + alpha-(5,6-dimethylbenzimidazolyl)deoxyadenosylcobamide
-
-
-
-
?
additional information
?
-
-
ATP:Cobalamin adenosyltransferases catalyze the transfer a 5'-deoxyadenosyl moiety from ATP to cob(I)alamin in the synthesis of the Co-C bond of coenzyme B12
-
-
?
additional information
?
-
-
MgATP and Cob(II)alamin binding sites, structure comparison overview
-
-
?
additional information
?
-
-
-
-
-
?
additional information
?
-
-
-
-
-
?
additional information
?
-
-
UTP, GTP, ITP are poor substrates
-
-
?
additional information
?
-
-
S-adenosylmethionine, vitamin B12r or B12a, AMP, ADP are no substrates
-
-
?
additional information
?
-
-
S-adenosylmethionine, vitamin B12r or B12a, AMP, ADP are no substrates
-
-
?
additional information
?
-
-
hydroxocobamides or cyanocobamides in which benzimidazole replaces 5,6-dimethylbenzimidazole, or cyanocobamide in which an adenine group replaces 5,6-dimethylbenzimidazole, can also act as substrates
-
-
?
additional information
?
-
-
the enzyme interacts with the methionine synthase reductase MSR, which catalyzes the reduction of cob(II)almin to cob(I)alamin, both enzymes activate each other, stoichiometry of the MSR-ATR system, overview
-
-
?
additional information
?
-
-
no activity with ADP and AMP
-
-
?
additional information
?
-
-
enzyme defects cause methylmalonic aciduria type B, regulation, overview
-
-
?
additional information
?
-
the enzyme catalyzes the final step in the conversion of cyanocobalamin, i.e. vitamin B12, to the essential human cofactor adenosylcobalamin, defects in the enzyme through mutations in the gene encoding the enzyme can result in the metabolic disorder known as methylmalonic aciduria, MMA
-
-
?
additional information
?
-
-
the enzyme catalyzes the final step in the conversion of cyanocobalamin, i.e. vitamin B12, to the essential human cofactor adenosylcobalamin, defects in the enzyme through mutations in the gene encoding the enzyme can result in the metabolic disorder known as methylmalonic aciduria, MMA
-
-
?
additional information
?
-
only two of the three active sites within the trimer contain the bound ATP substrate, twenty residues at the enzymes N-terminus become ordered upon binding of ATP to form an ATP-binding site and an extended cleft that likely binds cobalamin, cobalamin binding site structure involving residue R186, overview
-
-
?
additional information
?
-
-
only two of the three active sites within the trimer contain the bound ATP substrate, twenty residues at the enzymes N-terminus become ordered upon binding of ATP to form an ATP-binding site and an extended cleft that likely binds cobalamin, cobalamin binding site structure involving residue R186, overview
-
-
?
additional information
?
-
no in vitro activity by mutants R215K, R225K as well as K78Q, E84K, G87R, D90N, E91K, L92S, S94L, R186W, C189Y, R190C, R191W, E193K, R194G, F212S, S217R, L220P, and L223P
-
-
?
additional information
?
-
-
no in vitro activity by mutants R215K, R225K as well as K78Q, E84K, G87R, D90N, E91K, L92S, S94L, R186W, C189Y, R190C, R191W, E193K, R194G, F212S, S217R, L220P, and L223P
-
-
?
additional information
?
-
no mediation of base-off transition of adenosylcobalamin (key step of catalytic mechanism) by mutants D64G, F83S, G87R, D90N, E91K, L92S, S94L, C189Y, R191W, E193K, R194G, F212S, S217R, L220P, and L223P
-
-
?
additional information
?
-
-
no mediation of base-off transition of adenosylcobalamin (key step of catalytic mechanism) by mutants D64G, F83S, G87R, D90N, E91K, L92S, S94L, C189Y, R191W, E193K, R194G, F212S, S217R, L220P, and L223P
-
-
?
additional information
?
-
-
TTP does not serve as substrate
-
-
?
additional information
?
-
-
The PduO-type ATP:corrinoid adenosyltransferase catalyzes the transfer of the adenosyl-group of ATP to Co1+-cobalamin and Co1+-cobinamide substrates to synthesize adenosylcobalamin and adenosylcobinamide, respectively
-
-
?
additional information
?
-
-
to overcome the thermodynamically challenging Co2+ -> Co1+ reduction, the enzyme drastically weakens the axial ligand-Co2+ bond so as to generate effectively four-coordinate Co2+-corrinoid species, mechanism, overview. The entire hydrophobic pocket below the corrin ring, and not just residue F112, is critical for the removal of the axial ligand from the cobalt center of the Co2+-corrinoids. Large role of the ATP-induced active-site conformational changes with respect to the formation of 4c Co(II)Cbl
-
-
?
additional information
?
-
-
-
-
-
?
additional information
?
-
-
hydroxocobamides or cyanocobamides in which benzimidazole replaces 5,6-dimethylbenzimidazole, or cyanocobamide in which an adenine group replaces 5,6-dimethylbenzimidazole, can also act as substrates
-
-
?
additional information
?
-
-
-
-
?
additional information
?
-
-
-
-
-
?
additional information
?
-
cobyrinic acid is not a substrate
-
-
?
additional information
?
-
-
cobyrinic acid is not a substrate
-
-
?
additional information
?
-
-
crucial role of the 2'-OH group of the ribosyl moiety of ATP, the gamma-phosphate of ATP is critical for positioning the target for nucleophilic attack, differences in the base of the nucleotide have no effect on enzyme activity, 2'-deoxynucleotides fails to serve as substrates
-
-
?
additional information
?
-
-
EutT is necessary and sufficient for growth of a cobA eutT strain on ethanolamine as a carbon and energy or nitrogen source
-
-
?
additional information
?
-
-
the flavodoxin in vivo reducing agent that serves as the electron donor to the enzyme possesses a reduction potential that is considerably more positive than that of the Co2+/1+ couple of the corrinoid substrate, the enzyme overcomes this challenge by formation of an ATP-activated unique paramagnetic Co2+ corrinoid species by partial conversion, overview
-
-
?
additional information
?
-
-
enzyme-substrate interactions, intermediate/transition structures, spectroscopic mechanism analysis, computational modeling, overview
-
-
?
additional information
?
-
-
random or ordered ternary complex mechanism, the N-terminal domain is responsible for the catalytic activity and substrate binding and has similar biochemical properties and kinetic constants as the full-length enzyme
-
-
?
additional information
?
-
-
EuT substrate specificity, overview, reaction cycle involving EutT and CobA, overview
-
-
?
additional information
?
-
-
the enzyme can use free dihydroflavins to drive the adenosylation of cob(II)alamin
-
-
?
additional information
?
-
-
the enzyme is unable to remove the axial solvent ligand from cob(II)inamide and fails to accomplish the Co(II) to Co(I)rrinoid reduction under physiologically relevant conditions
-
-
?
additional information
?
-
in the active site, the corrin ring of Co(II)rrinoids is firmly locked in place by several amino acid side chains so as to facilitate the dissociation of the axial ligand
-
-
-
additional information
?
-
-
in the active site, the corrin ring of Co(II)rrinoids is firmly locked in place by several amino acid side chains so as to facilitate the dissociation of the axial ligand
-
-
-
additional information
?
-
-
the enzyme can use free dihydroflavins to drive the adenosylation of cob(II)alamin
-
-
?
additional information
?
-
EutT is necessary and sufficient for growth of a cobA eutT strain on ethanolamine as a carbon and energy or nitrogen source
-
-
?
additional information
?
-
-
GTP, CTP show only small activity as substrates
-
-
?
additional information
?
-
-
indications for cob(I)alamin-binding due to decrease in light absorbance by cob(I)alamin at 388 nm and conversion to a compound with absorbance maximum at 485 nm, different from adenosylcobalamin
-
-
?
additional information
?
-
indications for cob(I)alamin-binding due to decrease in light absorbance by cob(I)alamin at 388 nm and conversion to a compound with absorbance maximum at 485 nm, different from adenosylcobalamin
-
-
?
additional information
?
-
-
no ATP: cob(I)alamin adenosyltransferase activity detectable (anaerobic, 20 min, 80°C, pH 8, in presence of 1 mM titanium (III) citrate), no increase in light absorbance by presumably adenosylcobalamin at 525 nm
-
-
?
additional information
?
-
no ATP: cob(I)alamin adenosyltransferase activity detectable (anaerobic, 20 min, 80°C, pH 8, in presence of 1 mM titanium (III) citrate), no increase in light absorbance by presumably adenosylcobalamin at 525 nm
-
-
?
additional information
?
-
conserved residues are R119, R124, and E126
-
-
?
additional information
?
-
-
conserved residues are R119, R124, and E126
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
2 ATP + 2 cob(II)alamin + a reduced flavoprotein
2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
ATP + cob(I)alamin
triphosphate + adenosylcob(I)alamine
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
ATP + cob(I)alamin
triphosphate + adenosylcobalamin
ATP + cob(I)alamin
triphosphate + coenzyme B12
ATP + cob(I)yrinic acid a,c-diamide
triphosphate + adenosylcob(III)yrinic acid a,c-diamide
-
-
-
?
ATP + cobinamide
triphosphate + adenosylcobinamide
cob(I)alamin + ADP
adenosylcobalamin + diphosphate
-
-
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
cob(I)alamin + ATP + H2O + H+
adenosylcobalamin + diphosphate + phosphate
-
-
-
-
?
cob(I)alamin + CTP + H2O + H+
cytosylcobalamin + diphosphate + phosphate
-
-
-
-
?
cob(I)alamin + dATP + H2O + H+
deoxyadenosylcobalamin + diphosphate + phosphate
-
-
-
-
?
cob(I)alamin + GTP + H2O + H+
guanosylcobalamin + diphosphate + phosphate
-
low activity
-
-
?
cob(I)alamin + ITP
hypoxanthosylcobalamin + diphosphate + phosphate
-
low activity
-
-
?
cob(I)alamin + UTP
uranylcobalamin + diphosphate + phosphate
-
low activity
-
-
?
cob(I)inamide + ATP
adenosylcobinamide + triphosphate
-
-
-
-
?
additional information
?
-
2 ATP + 2 cob(II)alamin + a reduced flavoprotein
2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
-
-
-
?
2 ATP + 2 cob(II)alamin + a reduced flavoprotein
2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
-
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(I)alamine
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(I)alamine
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
-
adenosyltransferase enzymes lower the thermodynamic barrier of the Co2+ toCo+ reduction needed for the formation of the unique organometalic Co-C bond of adenosylcobalamin
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
-
adenosyltransferase enzymes lower the thermodynamic barrier of the Co2+ to Co+ reduction needed for the formation of the unique organometalic Co-C bond of adenosylcobalamin
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
final step in the conversion of vitamin B12 to coenzyme B12, the latter being required for degradation of 1,2-propanediol
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
last step in the corrinoid adenosylation pathway, EutT and CobA
-
-
ir
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
-
mechanism of adenosylcobalamin biosynthesis
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcob(III)alamin
last step in the corrinoid adenosylation pathway, EutT and CobA
-
-
ir
ATP + cob(I)alamin
triphosphate + adenosylcobalamin
-
polymorphic variants 239K and 239M, biosynthesis of adenosylcobalamin
-
-
?
ATP + cob(I)alamin
triphosphate + adenosylcobalamin
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
involved in vitamin B12 metabolism
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
-
-
-
?
ATP + cob(I)alamin
triphosphate + coenzyme B12
the main role of this enzyme is apparently the conversion of inactive cobalamins to adenosyl cobalamin for 1,2 propanediol degradation
-
-
?
ATP + cobinamide
triphosphate + adenosylcobinamide
-
-
-
-
?
ATP + cobinamide
triphosphate + adenosylcobinamide
-
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
adenosylcobalamin is a cofactor required by the methylmalonyl-CoA mutase
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
cobalamin assimilation and recycling pathway, overview, enzyme deficiency causes methylmalonic aciduria, MMA, is an autosomal recessive disease with symptoms that include ketoacidosis, lethargy, recurrent vomiting, dehydration, respiratory distress, muscular hypotonia and death due to methylmalonic acid levels that are up to 1000fold greater than normal, overview
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
adenosylation of the corrinoid ring of cob(I)alamin generates coenzyme B12, i.e. adenosylcobalamin or AdoCbl, an essential cofactor used by enzymes that catalyze intramolecular rearrangements, deaminations, dehydrations, reductions, and reductive dehalogenations
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
adenosylation of the corrinoid ring of cob(I)alamin generates coenzyme B12, i.e. adenosylcobalamin or AdoCbl, an essential cofactor used by enzymes that catalyze intramolecular rearrangements, deaminations, dehydrations, reductions, and reductive dehalogenations
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
-
-
-
?
cob(I)alamin + ATP
adenosylcobalamin + triphosphate
-
the enzyme is required to adenosylate de novo biosynthetic intermediates of adenosylcobalamin and to salvage incomplete and complete corrinoids from the environment of this bacterium
-
-
?
additional information
?
-
-
ATP:Cobalamin adenosyltransferases catalyze the transfer a 5'-deoxyadenosyl moiety from ATP to cob(I)alamin in the synthesis of the Co-C bond of coenzyme B12
-
-
?
additional information
?
-
-
the enzyme interacts with the methionine synthase reductase MSR, which catalyzes the reduction of cob(II)almin to cob(I)alamin, both enzymes activate each other, stoichiometry of the MSR-ATR system, overview
-
-
?
additional information
?
-
-
enzyme defects cause methylmalonic aciduria type B, regulation, overview
-
-
?
additional information
?
-
the enzyme catalyzes the final step in the conversion of cyanocobalamin, i.e. vitamin B12, to the essential human cofactor adenosylcobalamin, defects in the enzyme through mutations in the gene encoding the enzyme can result in the metabolic disorder known as methylmalonic aciduria, MMA
-
-
?
additional information
?
-
-
the enzyme catalyzes the final step in the conversion of cyanocobalamin, i.e. vitamin B12, to the essential human cofactor adenosylcobalamin, defects in the enzyme through mutations in the gene encoding the enzyme can result in the metabolic disorder known as methylmalonic aciduria, MMA
-
-
?
additional information
?
-
-
The PduO-type ATP:corrinoid adenosyltransferase catalyzes the transfer of the adenosyl-group of ATP to Co1+-cobalamin and Co1+-cobinamide substrates to synthesize adenosylcobalamin and adenosylcobinamide, respectively
-
-
?
additional information
?
-
-
EutT is necessary and sufficient for growth of a cobA eutT strain on ethanolamine as a carbon and energy or nitrogen source
-
-
?
additional information
?
-
-
the flavodoxin in vivo reducing agent that serves as the electron donor to the enzyme possesses a reduction potential that is considerably more positive than that of the Co2+/1+ couple of the corrinoid substrate, the enzyme overcomes this challenge by formation of an ATP-activated unique paramagnetic Co2+ corrinoid species by partial conversion, overview
-
-
?
additional information
?
-
-
EuT substrate specificity, overview, reaction cycle involving EutT and CobA, overview
-
-
?
additional information
?
-
EutT is necessary and sufficient for growth of a cobA eutT strain on ethanolamine as a carbon and energy or nitrogen source
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
0.27
Co2+
pH 8.0, 70°C, wild-type enzyme
0.00007 - 0.06
cob(I)alamin
0.000096
cob(I)inamide
-
-
0.0078 - 0.134
cob(II)alamin
0.0163
cob(II)inamide
wild type enzyme, pH and temperature not specified in the publication
0.11
Mg2+
pH 8.0, 70°C, wild-type enzyme
0.11
Mn2+
pH 8.0, 70°C, wild-type enzyme
additional information
additional information
-
0.0003
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91Y
0.0012
ATP
-
mutant F112Y, Co+ assay
0.0012
ATP
-
mutant F187A, Co+ assay
0.002
ATP
-
mutant F112W, Co+ assay
0.002
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91A/W93D
0.002
ATP
-
pH 8.0, 25°C, recombinant CobA mutant W93Y
0.0022
ATP
-
wild-type, Co+ assay
0.0026
ATP
-
mutant F112H, Co+ assay
0.0028
ATP
-
pH 8.0, 37°C
0.00287
ATP
mutant D218N, in presence of 0.05 mM cob(I)alamin
0.003
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91H
0.003
ATP
-
pH 8.0, 25°C, recombinant CobA mutant W93A
0.003
ATP
-
pH 8.0, 25°C, recombinant CobA mutant W93D
0.0031
ATP
-
mutant DELTAS183, Co+ assay
0.005
ATP
mutant T161I, in presence of 0.05 mM cob(I)alamin
0.005
ATP
-
wild-type, Co2+ assay
0.00523
ATP
mutant G97E, in presence of 0.05 mM cob(I)alamin
0.006
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91W/W93F
0.0063
ATP
-
recombinant enzyme variant 239K
0.0068
ATP
-
pH 8.0, 37°C, recombinant wild-type GST-tagged enzyme
0.0069
ATP
-
recombinant enzyme variant 239M
0.0069
ATP
native wild-type, in presence of 0.05 mM cob(I)alamin
0.007
ATP
-
pH 8.0, 25°C, recombinant wild-type CobA
0.00719
ATP
N-terminally octa-His tagged wild-type, in presence of 0.05 mM cob(I)alamin
0.00735
ATP
C-terminally octa-His tagged wild-type, in presence of 0.05 mM cob(I)alamin
0.0079
ATP
-
mutant F187A, Co2+ assay
0.00826
ATP
mutant K78R, in presence of 0.05 mM cob(I)alamin
0.009
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91A
0.009
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91W
0.0099
ATP
-
mutant DELTAS183, Co2+ assay
0.00997
ATP
mutant H183Y, in presence of 0.05 mM cob(I)alamin
0.01
ATP
-
pH 7.0, 37°C, recombinant enzyme
0.0104
ATP
-
mutant F112H, Co2+ assay
0.0124
ATP
mutant G63E, in presence of 0.05 mM cob(I)alamin
0.0141
ATP
mutant C119Y, in presence of 0.05 mM cob(I)alamin
0.0159
ATP
-
mutant F163A, Co+ assay
0.0172
ATP
mutant S68F, in presence of 0.05 mM cob(I)alamin
0.0198
ATP
-
pH 8.0, 37°C, recombinant full length wild-type enzyme
0.0203
ATP
mutant G97R, in presence of 0.05 mM cob(I)alamin
0.0205
ATP
mutant D64G, in presence of 0.05 mM cob(I)alamin
0.0254
ATP
wild type enzyme, pH and temperature not specified in the publication
0.026
ATP
-
pH 8.0, 25°C, recombinant CobA mutant W93H
0.03
ATP
-
pH 8.0, 25°C, recombinant CobA mutant W93F
0.0346
ATP
-
mutant F112A, Co+ assay
0.0459
ATP
mutant R76G, in presence of 0.05 mM cob(I)alamin
0.073
ATP
-
mutant F112W, Co2+ assay
0.0877
ATP
mutant S126L, in presence of 0.05 mM cob(I)alamin
0.094
ATP
-
mutant F112Y, Co2+ assay
0.0961
ATP
-
mutant F163A, Co2+ assay
0.11
ATP
pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+
0.215
ATP
mutant F83S, in presence of 0.05 mM cob(I)alamin
0.32
ATP
-
pH 8.0, 37°C, recombinant GST-tagged mutant R191W
0.33
ATP
pH 8.0, 70°C, mutant R124A, in presence of Mg2+
0.00007
cob(I)alamin
-
mutant DELTAS183, Co+ assay
0.00011
cob(I)alamin
-
mutant F187A, Co+ assay
0.00013
cob(I)alamin
-
wild-type, Co+ assay
0.00019
cob(I)alamin
-
mutant F163A, Co+ assay
0.00037
cob(I)alamin
-
pH 8.0, 37°C, recombinant wild-type GST-tagged enzyme
0.00055
cob(I)alamin
-
mutant F112Y, Co+ assay
0.00077
cob(I)alamin
-
mutant F112W, Co+ assay
0.001
cob(I)alamin
-
pH 8.0, 37°C
0.001
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91Y
0.0012
cob(I)alamin
-
recombinant enzyme variant 239K
0.00158
cob(I)alamin
N-terminally octa-His tagged wild-type, in presence of 0.5 mM ATP
0.0016
cob(I)alamin
-
recombinant enzyme variant 239M
0.0016
cob(I)alamin
C-terminally octa-His tagged wild-type, in presence of 0.5 mM ATP
0.0016
cob(I)alamin
native wild-type, in presence of 0.5 mM ATP
0.00172
cob(I)alamin
mutant D218N, in presence of 0.5 mM ATP
0.00186
cob(I)alamin
mutant S68F, in presence of 0.5 mM ATP
0.0019
cob(I)alamin
-
mutant F112A, Co+ assay
0.00194
cob(I)alamin
mutant D64G, in presence of 0.5 mM ATP
0.002
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant W93D
0.002
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant W93H
0.00224
cob(I)alamin
mutant G63E, in presence of 0.5 mM ATP
0.00225
cob(I)alamin
mutant K78R, in presence of 0.5 mM ATP
0.00252
cob(I)alamin
mutant G97E, in presence of 0.5 mM ATP
0.00271
cob(I)alamin
mutant T161I, in presence of 0.5 mM ATP
0.0028
cob(I)alamin
-
mutant F112H, Co+ assay
0.003
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91A/W93D
0.003
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91H
0.00303
cob(I)alamin
mutant G97R, in presence of 0.5 mM ATP
0.004
cob(I)alamin
-
at pH 7.0 and 37°C
0.004
cob(I)alamin
-
pH 8.0, 25°C, recombinant wild-type CobA
0.0041
cob(I)alamin
-
pH 7.0, 37°C, recombinant enzyme
0.0045
cob(I)alamin
-
pH 8.0, 37°C, recombinant full length wild-type enzyme
0.005
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91A
0.005
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91W
0.00513
cob(I)alamin
mutant C119Y, in presence of 0.5 mM ATP
0.0052
cob(I)alamin
-
pH 8.0, 37°C
0.00713
cob(I)alamin
mutant H183Y, in presence of 0.5 mM ATP
0.009
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91W/W93F
0.00909
cob(I)alamin
mutant R76G, in presence of 0.5 mM ATP
0.011
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant W93A
0.012
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant W93F
0.0143
cob(I)alamin
mutant S126L, in presence of 0.5 mM ATP
0.016
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant W93Y
0.0302
cob(I)alamin
mutant F83S, in presence of 0.5 mM ATP
0.06
cob(I)alamin
-
pH 8.0, 37°C, recombinant GST-tagged mutant R191W
0.0078
cob(II)alamin
-
wild-type, Co2+ assay
0.015
cob(II)alamin
-
mutant F187A, Co2+ assay
0.016
cob(II)alamin
-
mutant DELTAS183, Co2+ assay
0.028
cob(II)alamin
-
mutant F112W, Co2+ assay
0.034
cob(II)alamin
-
mutant F112Y, Co2+ assay
0.053
cob(II)alamin
-
mutant F112H, Co2+ assay
0.134
cob(II)alamin
-
mutant F163A, Co2+ assay
0.003
Cobalamin
pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+
0.004
Cobalamin
pH 8.0, 70°C, mutant R124A, in presence of Mg2+
0.01
cyanocob(I)alamin
-
-
0.01
cyanocob(I)alamin
-
pH 8.0, 37°C, i.e. vitamin B12, reduced form
0.14
dATP
-
0.14
dATP
pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+
0.003
hydroxocobalamin
-
0.003
hydroxocobalamin
+/- 0.0004
additional information
additional information
-
-
-
additional information
additional information
-
-
-
additional information
additional information
-
kinetics
-
additional information
additional information
-
kinetics
-
additional information
additional information
-
steady-state kinetics
-
additional information
additional information
elevated for either ATP or cobalamin or both substrates in case of mutants which show decreased adenosylcobalamin synthesis activity that can be partly corrected by increased hydroxycobalamin concentration
-
additional information
additional information
-
elevated for either ATP or cobalamin or both substrates in case of mutants which show decreased adenosylcobalamin synthesis activity that can be partly corrected by increased hydroxycobalamin concentration
-
additional information
additional information
large changes in KM for both substrates for mutants R76G, F83S and S126L
-
additional information
additional information
-
large changes in KM for both substrates for mutants R76G, F83S and S126L
-
additional information
additional information
wild-type kinetics by mutants G97E, C119Y, T161I, and H183Y
-
additional information
additional information
-
wild-type kinetics by mutants G97E, C119Y, T161I, and H183Y
-
additional information
additional information
-
wild-type: KM for CTP and UTP increased relative to ATP
-
additional information
additional information
-
wild-type: KM for GTP and ITP 530-13000fold increased relative to ATP
-
additional information
additional information
-
kinetics for wild-type and mutants enzymes from two different assay methods, overview
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
0.00041 - 297.6
cob(I)alamin
0.00067 - 0.26
cob(II)alamin
0.0077
cob(II)inamide
wild type enzyme, pH and temperature not specified in the publication
additional information
2'-deoxy-ATP
0.00038
ATP
-
mutant F112H, Co+ assay
0.0005
ATP
-
mutant F112H, Co2+ assay
0.0016
ATP
-
mutant F112Y, Co+ assay
0.0018
ATP
-
mutant F112W, Co+ assay
0.0023
ATP
-
mutant F112Y, Co2+ assay
0.0028
ATP
-
mutant F112W, Co2+ assay
0.0038
ATP
-
mutant S129A
0.0056
ATP
-
mutant F112A, Co+ assay
0.0062
ATP
-
mutant R128K
0.0067
ATP
wild type enzyme, pH and temperature not specified in the publication
0.008
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91A/W93D
0.0084
ATP
-
mutant R132K
0.01
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91W/W93F
0.01
ATP
-
pH 8.0, 25°C, recombinant CobA mutant W93F
0.014
ATP
-
mutant F187A, Co2+ assay
0.015
ATP
-
mutant F163A, Co+ assay
0.015
ATP
-
mutant F163A, Co2+ assay
0.017
ATP
-
mutant DELTAS183, Co2+ assay
0.02
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91A
0.02
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91H
0.021
ATP
-
mutant F187A, Co+ assay
0.023
ATP
-
mutant DELTAS183, Co+ assay
0.026
ATP
-
wild-type, Co+ assay
0.029
ATP
-
wild-type, Co2+ assay
0.03
ATP
-
pH 7.0, 37°C, recombinant enzyme
0.04
ATP
-
pH 8.0, 25°C, recombinant CobA mutant W93A
0.05
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91Y
0.05
ATP
-
pH 8.0, 25°C, recombinant CobA mutant W93D
0.07
ATP
-
pH 8.0, 25°C, recombinant CobA mutant W93H
0.07
ATP
mutant enzyme D110N, pH and temperature not specified in the publication
0.08
ATP
-
pH 8.0, 25°C, recombinant CobA mutant F91W
0.08
ATP
-
pH 8.0, 25°C, recombinant wild-type CobA
0.096
ATP
-
mutant D35E/R128K
0.1
ATP
-
pH 8.0, 25°C, recombinant CobA mutant W93Y
0.1
ATP
mutant enzyme R200K, pH and temperature not specified in the publication
0.14
ATP
pH 8.0, 70°C, mutant R124A, in presence of Mg2+
0.17
ATP
mutant enzyme K98Q, pH and temperature not specified in the publication
0.18
ATP
mutant enzyme E207Q, pH and temperature not specified in the publication
0.23
ATP
pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+
0.24
ATP
wild-type enzyme, pH and temperature not specified in the publication
0.27
ATP
mutant enzyme T88V, pH and temperature not specified in the publication
0.41
ATP
-
mutant D35N/R128K
9
ATP
-
mutant D35R/R128D, at subsaturating concentrations of 30 mM ATP and 0.02 mM cob(I)alamin
166.2
ATP
N-terminally octa-His tagged wild-type
297.6
ATP
C-terminally octa-His tagged wild-type
0.00041
cob(I)alamin
-
mutant R132K
0.00042
cob(I)alamin
-
mutant F112H, Co+ assay
0.00065
cob(I)alamin
-
mutant D35R/R128D, at subsaturating concentrations of ATP 30 mM ATP and 0.02 mM cob(I)alamin
0.0017
cob(I)alamin
-
mutant F112Y, Co+ assay
0.002
cob(I)alamin
-
mutant F112W, Co+ assay
0.0024
cob(I)alamin
-
mutant D35E/R128K
0.004
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91W/W93F
0.0066
cob(I)alamin
-
mutant F112A, Co+ assay
0.0068
cob(I)alamin
-
mutant D35N
0.009
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant W93F
0.009
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant W93Y
0.01
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91A/W93D
0.011
cob(I)alamin
-
pH 8.0, 37°C, recombinant GST-tagged mutant R191W
0.011
cob(I)alamin
-
mutant R128K
0.014
cob(I)alamin
-
mutant D35N/R128K
0.015
cob(I)alamin
-
mutant S129A
0.015
cob(I)alamin
-
mutant F163A, Co+ assay
0.018
cob(I)alamin
-
mutant F187A, Co+ assay
0.02
cob(I)alamin
-
mutant DELTAS183, Co+ assay
0.02
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91A
0.02
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91H
0.024
cob(I)alamin
-
wild-type, Co+ assay
0.037
cob(I)alamin
-
pH 8.0, 37°C, recombinant wild-type GST-tagged enzyme
0.05
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91Y
0.05
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant W93A
0.05
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant W93D
0.06
cob(I)alamin
-
pH 7.0, 37°C, recombinant enzyme
0.06
cob(I)alamin
-
at pH 7.0 and 37°C
0.08
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant F91W
0.08
cob(I)alamin
-
pH 8.0, 25°C, recombinant CobA mutant W93H
0.08
cob(I)alamin
-
pH 8.0, 25°C, recombinant wild-type CobA
19.2
cob(I)alamin
mutant K78R
29.4
cob(I)alamin
mutant G63E
31.8
cob(I)alamin
mutant D64G
34.8
cob(I)alamin
mutant S126L
43.8
cob(I)alamin
mutant D218N
46.8
cob(I)alamin
mutant S68F
48
cob(I)alamin
mutant G97R
124.2
cob(I)alamin
mutant F83S
137.4
cob(I)alamin
mutant T161I
154.8
cob(I)alamin
mutant R76G
166.2
cob(I)alamin
N-terminally octa-His tagged wild-type
169.8
cob(I)alamin
mutant G97E
169.8
cob(I)alamin
mutant H183Y
243.6
cob(I)alamin
mutant C119Y
285
cob(I)alamin
native wild-type
297.6
cob(I)alamin
C-terminally octa-His tagged wild-type
0.00067
cob(II)alamin
-
mutant F112H, Co2+ assay
0.0027
cob(II)alamin
-
mutant F112Y, Co2+ assay
0.0032
cob(II)alamin
-
mutant F112W, Co2+ assay
0.015
cob(II)alamin
-
mutant F187A, Co2+ assay
0.018
cob(II)alamin
-
mutant DELTAS183, Co2+ assay
0.027
cob(II)alamin
-
mutant F163A, Co2+ assay
0.038
cob(II)alamin
-
wild-type, Co2+ assay
0.05
cob(II)alamin
mutant enzyme D110N, pH and temperature not specified in the publication
0.06
cob(II)alamin
mutant enzyme R200K, pH and temperature not specified in the publication
0.18
cob(II)alamin
mutant enzyme K98Q, pH and temperature not specified in the publication
0.21
cob(II)alamin
wild-type enzyme, pH and temperature not specified in the publication
0.22
cob(II)alamin
mutant enzyme E207Q, pH and temperature not specified in the publication
0.26
cob(II)alamin
mutant enzyme T88V, pH and temperature not specified in the publication
0.11
Cobalamin
pH 8.0, 70°C, mutant R124A, in presence of Mg2+
0.18
Cobalamin
pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+
0.11
dATP
-
0.11
dATP
pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+
0.18
hydroxocobalamin
-
0.18
hydroxocobalamin
+/- 0.01
additional information
2'-deoxy-ATP
-
kcat/KM: 3300 1/M*s
additional information
ATP
C-terminally octa-His tagged wild-type, kcat/KM is 0.67 per microM and min
additional information
ATP
-
C-terminally octa-His tagged wild-type, kcat/KM is 0.67 per microM and min
additional information
ATP
-
kcat/KM: 12000 1/M*s
additional information
ATP
mutant C119Y, kcat/KM is 0.29 per microM and min
additional information
ATP
-
mutant C119Y, kcat/KM is 0.29 per microM and min
additional information
ATP
mutant D218N, kcat/KM is 0.25 per microM and min
additional information
ATP
-
mutant D218N, kcat/KM is 0.25 per microM and min
additional information
ATP
-
mutant D35E/R128K, kcat/KM: 23 1/M*s
additional information
ATP
-
mutant D35N, kcat/KM: 52 1/M*s
additional information
ATP
-
mutant D35N/R128K, kcat/KM: 32 1/M*s
additional information
ATP
-
mutant D35R/R128D, kcat/KM: 0.014 1/M*s, at subsaturating concentrations of 30 mM ATP and 0.02 mM cob(I)alamin
additional information
ATP
mutant D64G, kcat/KM is 0.026 per microM and min
additional information
ATP
-
mutant D64G, kcat/KM is 0.026 per microM and min
additional information
ATP
mutant F83S, kcat/KM is 0.010 per microM and min
additional information
ATP
-
mutant F83S, kcat/KM is 0.010 per microM and min
additional information
ATP
mutant G63E, kcat/KM is 0.039 per microM and min
additional information
ATP
-
mutant G63E, kcat/KM is 0.039 per microM and min
additional information
ATP
mutant G97E, kcat/KM is 0.46 per microM and min
additional information
ATP
-
mutant G97E, kcat/KM is 0.46 per microM and min
additional information
ATP
mutant G97R, kcat/KM is 0.039 per microM and min
additional information
ATP
-
mutant G97R, kcat/KM is 0.039 per microM and min
additional information
ATP
mutant H183Y, kcat/KM is 0.28 per microM and min
additional information
ATP
-
mutant H183Y, kcat/KM is 0.28 per microM and min
additional information
ATP
mutant K78R, kcat/KM is 0.039 per microM and min
additional information
ATP
-
mutant K78R, kcat/KM is 0.039 per microM and min
additional information
ATP
-
mutant R128K, kcat/KM: 1800 1/M*s
additional information
ATP
-
mutant R132K, kcat/KM: 39 1/M*s
additional information
ATP
mutant R76G, kcat/KM is 0.056 per microM and min
additional information
ATP
-
mutant R76G, kcat/KM is 0.056 per microM and min
additional information
ATP
mutant S126L, kcat/KM is 0.007 per microM and min
additional information
ATP
-
mutant S126L, kcat/KM is 0.007 per microM and min
additional information
ATP
-
mutant S129A, kcat/KM: 4500 1/M*s
additional information
ATP
mutant S68F, kcat/KM is 0.045 per microM and min
additional information
ATP
-
mutant S68F, kcat/KM is 0.045 per microM and min
additional information
ATP
mutant T161I, kcat/KM is 0.46 per microM and min
additional information
ATP
-
mutant T161I, kcat/KM is 0.46 per microM and min
additional information
ATP
N-terminally octa-His tagged wild-type, kcat/KM is 0.39 per microM and min
additional information
ATP
-
N-terminally octa-His tagged wild-type, kcat/KM is 0.39 per microM and min
additional information
ATP
native wild-type, kcat/KM is 0.69 per microM and min
additional information
ATP
-
native wild-type, kcat/KM is 0.69 per microM and min
additional information
cob(I)alamin
C-terminally octa-His tagged wild-type, kcat/KM is 3.10 per microM and min
additional information
cob(I)alamin
-
C-terminally octa-His tagged wild-type, kcat/KM is 3.10 per microM and min
additional information
cob(I)alamin
-
kcat/KM: 180000 1/M*s
additional information
cob(I)alamin
mutant C119Y, kcat/KM is 0.79 per microM and min
additional information
cob(I)alamin
-
mutant C119Y, kcat/KM is 0.79 per microM and min
additional information
cob(I)alamin
mutant D218N, kcat/KM is 0.42 per microM and min
additional information
cob(I)alamin
-
mutant D218N, kcat/KM is 0.42 per microM and min
additional information
cob(I)alamin
-
mutant D35E/R128K, kcat/KM: 900 1/M*s
additional information
cob(I)alamin
-
mutant D35N, kcat/KM: 2800 1/M*s
additional information
cob(I)alamin
-
mutant D35N/R128K, kcat/KM: 3800 1/M*s
additional information
cob(I)alamin
-
mutant D35R/R128D, kcat/KM: 5.2 1/M*s, at subsaturating concentrations of 30 mM ATP and 0.02 mM cob(I)alamin
additional information
cob(I)alamin
mutant D64G, kcat/KM is 0.27 per microM and min
additional information
cob(I)alamin
-
mutant D64G, kcat/KM is 0.27 per microM and min
additional information
cob(I)alamin
mutant F83S, kcat/KM is 0.07 per microM and min
additional information
cob(I)alamin
-
mutant F83S, kcat/KM is 0.07 per microM and min
additional information
cob(I)alamin
mutant G63E, kcat/KM is 0.22 per microM and min
additional information
cob(I)alamin
-
mutant G63E, kcat/KM is 0.22 per microM and min
additional information
cob(I)alamin
mutant G97E, kcat/KM is 0.94 per microM and min
additional information
cob(I)alamin
-
mutant G97E, kcat/KM is 0.94 per microM and min
additional information
cob(I)alamin
mutant G97R, kcat/KM is 0.26 per microM and min
additional information
cob(I)alamin
-
mutant G97R, kcat/KM is 0.26 per microM and min
additional information
cob(I)alamin
mutant H183Y, kcat/KM is 0.40 per microM and min
additional information
cob(I)alamin
-
mutant H183Y, kcat/KM is 0.40 per microM and min
additional information
cob(I)alamin
mutant K78R, kcat/KM is 0.14 per microM and min
additional information
cob(I)alamin
-
mutant K78R, kcat/KM is 0.14 per microM and min
additional information
cob(I)alamin
-
mutant R128K, kcat/KM: 12000 1/M*s
additional information
cob(I)alamin
-
mutant R132K, kcat/KM: 55 1/M*s
additional information
cob(I)alamin
mutant R76G, kcat/KM is 0.28 per microM and min
additional information
cob(I)alamin
-
mutant R76G, kcat/KM is 0.28 per microM and min
additional information
cob(I)alamin
mutant S126L, kcat/KM is 0.04 per microM and min
additional information
cob(I)alamin
-
mutant S126L, kcat/KM is 0.04 per microM and min
additional information
cob(I)alamin
-
mutant S129A, kcat/KM: 65000 1/M*s
additional information
cob(I)alamin
mutant S68F, kcat/KM is 0.42 per microM and min
additional information
cob(I)alamin
-
mutant S68F, kcat/KM is 0.42 per microM and min
additional information
cob(I)alamin
mutant T161I, kcat/KM is 0.85 per microM and min
additional information
cob(I)alamin
-
mutant T161I, kcat/KM is 0.85 per microM and min
additional information
cob(I)alamin
N-terminally octa-His tagged wild-type, kcat/KM is 1.75 per microM and min
additional information
cob(I)alamin
-
N-terminally octa-His tagged wild-type, kcat/KM is 1.75 per microM and min
additional information
cob(I)alamin
native wild-type, kcat/KM is 2.97 per microM and min
additional information
cob(I)alamin
-
native wild-type, kcat/KM is 2.97 per microM and min
additional information
CTP
-
kcat/KM: 0.38 1/M*s
additional information
GTP
-
kcat/KM: 18 1/M*s
additional information
ITP
-
kcat/KM: 0.65 1/M*s
additional information
additional information
-
recombinant truncated enzyme versions
-
additional information
additional information
-
adenosylation of cobalamin and cobinamide (lacking dimethylbenzimidazole moiety) at similar rates
-
additional information
additional information
kcat of N-terminal octa-His tagged enzyme is 58% of native or C-terminal tagged enzyme possibly due to interference of N-terminal tag with base-off transition of adenosylcobalamin by the enzyme
-
additional information
additional information
-
kcat of N-terminal octa-His tagged enzyme is 58% of native or C-terminal tagged enzyme possibly due to interference of N-terminal tag with base-off transition of adenosylcobalamin by the enzyme
-
additional information
additional information
mutations in conserved regions E84-S94 and R186-R194 lead to abolished enzymatic activity and support their implication in the enzymes active site
-
additional information
additional information
-
mutations in conserved regions E84-S94 and R186-R194 lead to abolished enzymatic activity and support their implication in the enzymes active site
-
additional information
additional information
-
wild-type, decreased kcat for CTP and UTP relative to ATP
-
additional information
additional information
-
wild-type, kcat for GTP and ITP is not decreased relative to ATP
-
additional information
UTP
-
kcat/KM: 0.048 1/M*s
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apoenzyme and in complex with Mg2+/ATP, C-centred orthorhombic space group C222(1), unit cell parameters: a: 64.93 A, b: 137.08 A, c: 158.55 A, alpha, beta, gamma: 90°, one trimer in the asymmetric unit, sitting-drop combined with hanging-drop vapour-diffusion method: 13 mg/ml protein solution, precipitants: 1.55-1.6 M ammonium sulfate, 9-10% (v/v) dioxane (pH 6.5), for complex: 4 mM ATP and 4 mM Mg2+
-
purified recombinant PduO in complex with ATP, hanging drop vapor diffusion method, mixing o 0.001 ml of 13 mg/ml protein solution, with or without 4 mM ATP and 4 mM MgCl2, with 0.001 ml of optimized reservoir solution containing 100 mM MES, pH 6.5, 1.52 M ammonium sulfate, 9% v/v dioxane, followed by equilibration over 0.5 ml of the mother liquor, the cryoprotection solution contains 50 mM MES, pH 6.5, 0.76 M ammonium sulfate, 4.5% v/v dioxane, and 1.7 M sodium malonate, pH 7.0, X-ray diffraction structure determination and analysis
-
native (PDB: 2ZHY, with ordered N-terminal loop and formed active site) and in complex with ATP (PDB: 2ZHZ, substrate-binding cleft is widened and the N-terminal loop swung out and conformational shift of Arg129 side chain upon ATP-binding, similar structure to human and Lactobacillus reuteri PduO except for interaction of NH2 nitrogen atom of Arg11 with gamma-phosphate of ATP), five alpha helix bundle, monomeric subunits almost identical conformations in both structures, crystals: orthothrombic space group C222(1), three monomers in the asymmetric unit, unit cell parameters: a: 52.55/53.91, b: 148.98/148.17, c: 157.35/158.10, microcrystals (from sitting-drop vapour-diffusion at 22°C) scaled up by hanging-drop vapour diffusion, reservoir solution (pH5.7, 22% (w/v) isopropanol, 12% (v/v) PEG4000), PduO-MgATP complex: ATP soaked into native PduO crystals, molecular replacement using PDB: 2G2D as model
mutants D35N (without tag) in complex with ATP and cob(II)alamin and R132K (without tag) in complex with ATP, thin plate crystals, space group P6(3), two monomers in the asymmetric unit, unit cell parameters: a, b: 65A, c: 169A, beta: 90°; vapour-diffusion under anoxic conditions, protein solution (15 mg/ml, containing ATP, hydroxycobalamin and a reducing system of NADH, FMN, and flavodoxin reductase), reservoir solution (incl. 14-16% PEG8000, pH6)
-
purified recombinant His-tagged enzyme in complex with its substrates, hanging drop vapour diffusion method, 20°C, 0.004 ml of 20 mg/ml protein in 10 mM Tris-HCl, pH 8.0, is mixed with 0.004 ml od precipitant solution containing 0.1 M HEPES, pH 8.5, 1.1 M ammonium sulfate, 2 mM ATP, 55 mM MgCl2, 165 mM NaCl, and 15 mM cob(I)alamin, 7 days, X-ray diffraction structure determination and analysis at 1.68 A resolution
-
trimer of three independent five-helix bundles, active sites at the interface between adjacent monomers, no significant structural changes accompany catalysis, precatalytic complex with ATP: cob(II)alamin (PDB: 3CI1, four-coordinate, base-off cob(II)alamin intermediate, enzyme with fully ordered six C-terminal residues and potassium ion in active site), complex with tripolyphosphate: adenosylcobalamin (PDB: 3CI3, partially occupied with five-coordinate adenosylcobalamin), precatalytic complex with ATP: cob(II)inamide (PDB: 3CI4, cob(II)inamide-binding structurally indistinguishable from cob(II)alamin-binding), binding of cobalamin and cobinamide (lacking dimethylbenzimidazole moiety) in identical positions and orientation, space group R3, one molecule in asymmetric unit, unit cell parameters: a: 67.8-68, b: 67.8-68, c: 110.9-111.3, beta: 90°, molecular replacement using PDB: 2NT8 as model; vapour-diffusion with tag-cleaved protein solution (18-22 mg/ml, in presence of hydroxycobalamin and/or adenosylcobalamin or dicyanocobinamide, ATP etc.) and reservoir solution (10-13% (w/v) PEG 8000, pH 6), cubic crystals, crystallisation under anoxic conditions in presence of flavin-dependent reducing system
-
purified recombinant CobA in complex with ATP, four-coordinate cobalamin, and five-coordinate cobalamin, hanging drop vapour diffusion method, 0.002 ml of 10 mg/ml CobA protein in 20 mM Tris-HCl, pH 8.0, 20 mM NADH, 3 mM ATP, 4.5 mM MgCl2, and 2 mM HOCbl, is mixed with 0.002 ml of well solution containing 100 mM MES, pH 6.0, 320 mM NaCl, and 19.6% w/v PEG4000, X-ray diffraction structure determination and analysis at 1.95 A resolution
-
sitting drop vapour diffusion method. The structure of PduOC co-crystallized with heme is solved (1.9 A resolution) showing an octameric assembly with four heme moieities
3fold symmetric trimer of five counterclockwise helix bundles, one molecule in asymmetric unit, polypropylene glycol 400 molecule captured in putative active site (positively charged, important residues: Asp32, Arg118), crystals of selenomethionine derivative: space group P2(1)3, unit cell parameters: a, b, c: 84.67 A, hanging-drop vapour-diffusion method: protein solution (15 mg/ml) + reservoir solution (pH 8.1, 2.5 M ammonium sulphate, 2% polypropylene glycol 400)
hanging-drop vapor diffusion method
sparse matrix method at 20°C
trimer with noncrystallographic 3fold symmetry in the asymmetric unit, consistent of five helix bundles, identical topology to ST1454 but less ion pairs around the putative active site, crystals: space group P4(1)2(1)2, unit cell parameters: a, b: 117.58 A, c: 79.05 A, hanging-drop vapour-diffusion method: protein solution (21 mg/ml) + reservoir solution (pH6.8, 0.5 M ammonium sulfate, 2% polypropylene glycol 400), molecular replacement
purified recombinant wild-type and selenomethionine-labeled enzymes, crystal growth from 0.4 M ammonium phosphate, 4% methyl-pentanediol, 5% glycerol, at 20°C, X-ray diffraction structure determination and analysis at 1.5-1.9 A resolution
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C119Y
wild-type kinetics, decreased adenosylcobalamin production in vivo, impaired protein folding leads to degradation and, thus, low expression, no rescue of ATR-deficient Salmonella strain BE620
C189Y
inactive in vitro (10fold excess of substrate compared to standard), decreased adenosylcobalamin production in vivo, impaired protein folding leads to degradation and, thus, low expression (but can be purified), no rescue of ATR-deficient Salmonella strain BE620
D218N
substantially reduced Vmax, decreased adenosylcobalamin production in vivo partly corrected by increased hydroxycobalamin concentration, part of proposed active site role in ATP/cobalamin binding, no rescue of ATR-deficient Salmonella strain BE620
D64G
substantially reduced Vmax, decreased adenosylcobalamin production in vivo partly corrected by increased hydroxycobalamin concentration, part of proposed active site, role in ATP/cobalamin binding, no rescue of ATR-deficient Salmonella strain BE620
D90N
inactive in vitro (10fold excess of substrate compared to standard)
E84K
inactive in vitro (10fold excess of substrate compared to standard)
E91K
inactive in vitro (10fold excess of substrate compared to standard)
F212S
inactive in vitro (10fold excess of substrate compared to standard)
F83S
large change in KM for ATP and cob(I)alamin, F83 has direct contact with ATP
G63E
substantially reduced Vmax, decreased adenosylcobalamin production in vivo partly corrected by increased hydroxycobalamin concentration, role in ATP/cobalamin binding, no rescue of ATR-deficient Salmonella strain BE620
G87R
inactive in vitro (10fold excess of substrate compared to standard)
G97E
wild-type kinetics, mutation distant from proposed active site, no rescue of ATR-deficient Salmonella strain BE620 possibly due to impaired reduction of cob(II)alamin to cob(I)alamin, expressed at wild-type levels
G97R
substantially reduced Vmax, mutation distant from proposed active site, no rescue of ATR-deficient Salmonella strain BE620
H183Y
wild-type kinetics, mutation distant from proposed active site, no rescue of ATR-deficient Salmonella strain BE620 possibly due to impaired reduction of cob(II)alamin to cob(I)alamin, expressed at wild-type levels
K78Q
inactive in vitro (10fold excess of substrate compared to standard)
K78R
substantially reduced Vmax, decreased adenosylcobalamin production in vivo partly corrected by increased hydroxycobalamin concentration, role in ATP/cobalamin binding, no rescue of ATR-deficient Salmonella strain BE620
L220P
inactive in vitro (10fold excess of substrate compared to standard)
L223P
inactive in vitro (10fold excess of substrate compared to standard)
L92S
inactive in vitro (10fold excess of substrate compared to standard)
R190C
inactive in vitro (10fold excess of substrate compared to standard), decreased adenosylcobalamin production in vivo, impaired protein folding leads to degradation and, thus, low expression (but can be purified), no rescue of ATR-deficient Salmonella strain BE620, conserved residue, mutation found in methylmalonic aciduria patients
R190H
-
catalytically inactive patient mutation leading to the inherited disorder methylmalonic aciduria. Mutant is examined using intrinsic fluorescence quenching of MMAB as a measure of ligand-binding. R190H and R186W significantly disrupt the affinity between MMAB and adenosylcobalmin. Arg 186 and Arg-190 may be critical for the transfer of the 5'-deoxyadenosyl group from ATP to cob(I)alamin, possibly by contributing to the precise positioning of the two substrates to permit catalysis to occur
R191W
inactive in vitro (10fold excess of substrate compared to standard), decreased adenosylcobalamin production in vivo, impaired protein folding leads to degradation and, thus, low expression (but can be purified), no rescue of ATR-deficient Salmonella strain BE620, conserved residue, mutation found in methylmalonic aciduria patients
R191W/A135T
-
site-directed mutagenesis, the mutant enzyme shows 30% reduced activity compared to the wild-type enzyme
R194G
inactive in vitro (10fold excess of substrate compared to standard)
R215K
inactive in vitro (10fold excess of substrate compared to standard), lack of activity in vitro, decreased adenosylcobalamin production in vivo partly corrected by increased hydroxycobalamin concentration, part of proposed active site, role in ATP/cobalamin binding, no rescue of ATR-deficient Salmonella strain BE620
R225K
lack of activity in vitro, decreased adenosylcobalamin production in vivo partly corrected by increased hydroxycobalamin concentration, part of proposed active site, role in ATP/cobalamin binding, no rescue of ATR-deficient Salmonella strain BE620
R76G
large change in KM for ATP and cob(I)alamin, decreased adenosylcobalamin production in vivo partly corrected by increased hydroxycobalamin concentration, part of proposed active site, role in ATP/cobalamin binding, no rescue of ATR-deficient Salmonella strain BE620
S126L
large change in KM for ATP and cob(I)alamin, decreased adenosylcobalamin production in vivo partly corrected by increased hydroxycobalamin concentration, part of proposed active site, role in ATP/cobalamin binding, no rescue of ATR-deficient Salmonella strain BE620
S217R
inactive in vitro (10fold excess of substrate compared to standard)
S68F
substantially reduced Vmax, residue S68 has role in ATP-binding
S94L
inactive in vitro (10fold excess of substrate compared to standard)
T161I
wild-type kinetics, decreased adenosylcobalamin production in vivo but rescues ATR-deficient Salmonella strain BE620 possibly due to impaired reduction of cob(II)alamin to cob(I)alamin, expressed at wild-type levels
D35E/R128K
-
reduced activity to lower extent than mutation R128K alone
D35N/R128K
-
similar kinetics as mutation D35N alone
D35R/R128D
-
reciprocal mutation to D35/R128, very high KM values did not allow for kinetic analyses at saturating substrate concentrations
DELTAS183
-
Km (mM) (Co+ assay): 0.0031 (ATP), 0.00007 (cob(I)alamin), kcat (1/sec) (Co+ assay): 0.023 (ATP), 0.02 (cob(I)alamin), Km (mM) (Co2+ assay): 0.0099 (ATP), 0.0163 (cob(II)alamin), kcat (1/sec) (Co+ assay): 0.017 (ATP), 0.018 (cob(II)alamin)
F112W
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Km (mM) (Co+ assay): 0.002 (ATP), 0.00077 (cob(I)alamin), kcat (1/sec) (Co+ assay): 0.0018 (ATP), 0.002 (cob(I)alamin), Km (mM) (Co2+ assay): 0.0073 (ATP), 0.028 (cob(II)alamin), kcat (1/sec) (Co+ assay): 0.0028 (ATP), 0.0032 (cob(II)alamin)
F112Y
-
Km (mM) (Co+ assay): 0.0012 (ATP), 0.00055 (cob(I)alamin), kcat (1/sec) (Co+ assay): 0.0016 (ATP), 0.0017 (cob(I)alamin), Km (mM) (Co2+ assay): 0.0094 (ATP), 0.034 (cob(II)alamin), kcat (1/sec) (Co+ assay): 0.0023 (ATP), 0.0027 (cob(II)alamin)
F163A
-
Km (mM) (Co+ assay): 0.0159 (ATP), 0.00019 (cob(I)alamin), kcat (1/sec) (Co+ assay): 0.015 (ATP), 0.015 (cob(I)alamin), Km (mM) (Co2+ assay): 0.096 (ATP), 0.134 (cob(II)alamin), kcat (1/sec) (Co+ assay): 0.015 (ATP), 0.027 (cob(II)alamin)
S129A
-
mildly affected kcat and KM for both substrated
S159A
-
site-directed mutagenesis
V186A
-
site-directed mutagenesis
D110R
no activity detected with the mutant enzyme
D110R/R100D
no activity detected with the mutant enzyme
N231L
no activity detected with the mutant enzyme
R200D
no activity detected with the mutant enzyme
R204K
no activity detected with the mutant enzyme
D110R
-
no activity detected with the mutant enzyme
-
D110R/R100D
-
no activity detected with the mutant enzyme
-
N231L
-
no activity detected with the mutant enzyme
-
R200D
-
no activity detected with the mutant enzyme
-
R204K
-
no activity detected with the mutant enzyme
-
A134L
-
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
F91A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
F91A,W93A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
F91D
-
site-directed mutagenesis, the mutant is inactive
F91H
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
F91W,W93F
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
F91W/W93F
in this mutant the yield of 4c cob(II)alamin species is reduced more than 10fold from that achieved by the wild type enzyme
H67A
-
the mutant shows 100fold reduced catalytic efficiency compared to the wild type enzyme
R165A
-
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
R165E
-
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
R98A
-
site-directed mutagenesis, the mutant shows 80% reduced activity compared to the wild-type enzyme
R98E
-
site-directed mutagenesis, the mutant shows about 30% reduced activity compared to the wild-type enzyme
R98E/R165E
-
site-directed mutagenesis, the mutant shows 85% reduced activity compared to the wild-type enzyme
R9A
-
site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type enzyme
R9E
-
site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type enzyme
R9E/R165E
-
site-directed mutagenesis, the mutant shows 30% reduced activity compared to the wild-type enzyme
R9E/R98E
-
site-directed mutagenesis, the mutant shows 60% reduced activity compared to the wild-type enzyme
R9E/R98E/R165E
-
site-directed mutagenesis, the mutant shows 25% reduced activity compared to the wild-type enzyme
W93D
-
site-directed mutagenesis, the mutant shows the same activity as the wild-type enzyme
W93Y
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
C79A
-
the mutant shows 1 order of magnitude reduced catalytic efficiency compared to the wild type enzyme
-
C83A
-
the mutant shows 30fold reduced catalytic efficiency compared to the wild type enzyme
-
H67A
-
the mutant shows 100fold reduced catalytic efficiency compared to the wild type enzyme
-
E193K
-
site-directed mutagenesis, the mutant enzyme is not expressed
E193K
inactive in vitro (10fold excess of substrate compared to standard), conserved residue, mutation found in methylmalonic aciduria patients
R186W
inactive in vitro (10fold excess of substrate compared to standard), decreased adenosylcobalamin production in vivo, impaired protein folding leads to degradation and, thus, low expression (but can be purified), no rescue of ATR-deficient Salmonella strain BE620, conserved residue, mutation found in methylmalonic aciduria patients
R186W
-
catalytically inactive patient mutation leading to the inherited disorder methylmalonic aciduria. Mutant is examined using intrinsic fluorescence quenching of MMAB as a measure of ligand-binding. R190H and R186W significantly disrupt the affinity between MMAB and adenosylcobalmin. Arg 186 and Arg-190 may be critical for the transfer of the 5'-deoxyadenosyl group from ATP to cob(I)alamin, possibly by contributing to the precise positioning of the two substrates to permit catalysis to occur
D35N
-
site-directed mutagenesis
D35N
-
230fold decrease in kcat/KM (ATP) most likely due to disruption of salt bridge with residue R128 as observed in crystal structure
F112A
-
site-directed mutagenesis
F112A
-
Km (mM) (Co+ assay): 0.0346 (ATP), 0.0019 (cob(I)alamin), kcat (1/sec) (Co+ assay): 0.0056 (ATP), 0.006 (cob(I)alamin). Mutant F112A shows no activity in the Co2+ assay. The crystal structure of mutant F112A reveal that cob(II)alamin binds to the active site as a five-coordinate species with 5,6-dimethylbenzimidazole serving as a fifth axial ligand. In the Co+ assay [adenosylation of cob(I)alamin], mutant F112A is catalytically competent and displays only a slight decrease in kcat, supposing that in the absence of a four-coordinate cob(II)alamin species, the enzyme is inactive
F112A
-
mutation of the conserved Phe-112 in the active site of PduO is critical for the displacement of cob(II)alamin and leads to an inactive enzyme
F112H
-
site-directed mutagenesis
F112H
-
Km (mM) (Co+ assay): 0.0026 (ATP), 0.0028 (cob(I)alamin), kcat (1/sec) (Co+ assay): 0.00038 (ATP), 0.0042 (cob(I)alamin), Km (mM) (Co2+ assay): 0.01 (ATP), 0.053 (cob(II)alamin), kcat (1/sec) (Co+ assay): 0.00059 (ATP), 0.00067 (cob(II)alamin). The crystal structure of an mutantF112H variant show a 5,6-dimethylbenzimidazole-off/His-on interaction between the corrinoid and the enzyme, whose catalytic efficiency is 4 orders of magnitude lower than that of the wild-type protein. The analysis of the kinetic parameters of mutantF112H suggests that the F112H substitution negatively impacts product release
F187A
-
site-directed mutagenesis
F187A
-
Km (mM) (Co+ assay): 0.0012 (ATP), 0.00011 (cob(I)alamin), kcat (1/sec) (Co+ assay): 0.021 (ATP), 0.018 (cob(I)alamin), Km (mM) (Co2+ assay): 0.0079 (ATP), 0.0158 (cob(II)alamin), kcat (1/sec) (Co+ assay): 0.014 (ATP), 0.015 (cob(II)alamin)
R128K
-
site-directed mutagenesis
R128K
-
kinetics similar to wild-type, R128 conserved among PduO-type ACAs, R128 and R128K build salt bridge to residue D35 of adjacent unit, mutation R128W most common in methylmalonic aciduria patients
R132K
-
site-directed mutagenesis
R132K
-
60-80fold decrease in kcat with respect to both substrates, 300fold decrease in kcat/KM (ATP), 3000fold decrease in kcat/KM (cob(I)alamin), identical position of ATP in the crystal structure compared to wild-type, no cob(I)alamin detectable in crystal structure
C79A
-
site-directed mutagenesis of EutT, the mutant shows highly reduced activity compared to the wild-type enzyme
C79A
-
the mutant shows 1 order of magnitude reduced catalytic efficiency compared to the wild type enzyme
C80A
-
inactive
C80A
-
site-directed mutagenesis of EutT, the mutant shows 99% reduced activity compared to the wild-type enzyme
C83A
-
site-directed mutagenesis of EutT, the mutant shows 99% reduced activity compared to the wild-type enzyme
C83A
-
the mutant shows 30fold reduced catalytic efficiency compared to the wild type enzyme
F91W
-
site-directed mutagenesis, the mutant shows the same activity as the wild-type enzyme
F91W
with cob(II)alamin the mutant shows catalytic activity diminished relative to that of wild type enzyme, while the activity with cob(I)alamin is largely retained
F91Y
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
F91Y
with cob(II)alamin the mutant shows catalytic activity 5fold increased relative to that of wild type enzyme
W93A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
W93A
the mutation completely abolishes the catalytic activity with cob(II)alamin while modest activity is retained with cob(I)alamin
W93F
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
W93F
the mutant shows wild type activity
W93H
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
W93H
the mutation inhibits the formation of 4c cob(II)alamin
E126A
inactive mutant
E126A
no catalytic activity
E126A
site-directed mutagenesis, inactive mutant
E126K
inactive mutant
E126K
no catalytic activity
E126K
site-directed mutagenesis, inactive mutant
R119A
inactive mutant
R119A
no catalytic activity
R119A
site-directed mutagenesis, inactive mutant
R119W
inactive mutant
R119W
no catalytic activity
R119W
site-directed mutagenesis, inactive mutant
R124A
reduced activity
R124A
about half as active as wild-type
R124A
site-directed mutagenesis, about 40% reduced activity compared to the wild-type enzyme
R124F
reduced activity
R124F
decreased kcat and increased KM
R124F
site-directed mutagenesis, about 95% reduced activity compared to the wild-type enzyme
R124K
inactive mutant
R124K
no catalytic activity
R124K
site-directed mutagenesis, inactive mutant
R124W
inactive mutant
R124W
no catalytic activity
R124W
site-directed mutagenesis, inactive mutant
additional information
-
construction of transgenic C57/Bl6 mice by expression of the human enzyme using an adeno-associated virus vector with primer pairs specific to the cytomegalovirus enhancer/chicken beta-actin, CBAT, promoter, quantitative and semiquantitative expression analysis in murine liver, overview
additional information
enzyme mutations at residues Gly97, Ser174, Arg186, Arg190, Arg191, Glu193, and Gln234 can result in the metabolic disorder known as methylmalonic aciduria, MMA, overview
additional information
-
enzyme mutations at residues Gly97, Ser174, Arg186, Arg190, Arg191, Glu193, and Gln234 can result in the metabolic disorder known as methylmalonic aciduria, MMA, overview
additional information
-
enzyme expression restores coenzyme B12 synthesis in a Salmonella enterica strain lacking the housekeeping CobA enzyme
additional information
-
construction of several truncated mutants, the activity decreases with truncation degree, remaining residues 1-90 are inactive, overview
additional information
-
truncation of the 9-amino acid N-terminal helix of CobA reduces its FldA-dependent cobalamin adenosyltransferase activity by 97.4%, but shows 4fold higher specific activity than the wild-type enzyme when cob(I)alamin is generated chemically in situ, e.g. by use of KBH4, residues Arg9 and Arg165 of CobA are critical for FldA-dependent adenosylation but are catalytically as competent as the wild-type protein when cob(I)alamin i provided as substrate, overview
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Vitols, E.; Walker, G.A.; Huennekens, F.M.
Enzymatic conversion of vitamin B-12s to a cobamide coenzyme, alpha-(5,6-dimethylbenzimidazolyl)deoxyadenosylcobamide (adenosyl-B-12)
J. Biol. Chem.
241
1455-1461
1966
Clostridium tetanomorphum
brenda
Mudd, S.H.
The adenosyltransferases
The Enzymes, 3rd. Ed. (Boyer P. D. ed. )
8
121-154
1973
Clostridium tetanomorphum, Propionibacterium freudenreichii subsp. shermanii
-
brenda
Beck, W.S.
Ribosome-associated vitamin B12s adenosylating enzyme of Lactobacillus leichmannii
Methods Enzymol.
67
41-56
1980
Clostridium tetanomorphum, Lactobacillus delbrueckii, Lactobacillus leichmannii, Propionibacterium freudenreichii subsp. shermanii
brenda
Parry, R.J.; Ostrander, J.M.; Arzu, I.Y.
Studies of enzyme stereochemistry. Elucidation of the stereochemistry of the reaction catalyzed by cob(I)alamin adenosyltransferase
J. Am. Chem. Soc.
107
2190-2191
1985
Clostridium tetanomorphum
-
brenda
Debussche, L.; Couder, M.; Thibaut, D.; Cameron, B.; Crouzet, J.; Blanche, F.
Purification and partial characterization of Cob(I)alamin adenosyltransferase from Pseudomonas denitrificans
J. Bacteriol.
173
6300-6302
1991
Pseudomonas denitrificans (nom. rej.) (P29930), Pseudomonas denitrificans (nom. rej.)
brenda
Crouzet, J.; Levy-Schil, S.; Cameron, B.; Cauchois, L.; Rigault, S.; Rouyez, M.C.; Blanche, F.; Debussche, L.; Thibaut, D.
Nucleotide sequence and genetic analysis of a 13.1-kilobase-pair Pseudomonas denitrificans DNA fragment containing five cob genes and identification of structural genes encoding Cob(I)alamin adenosyltransferase, cobyric acid synthase, and bifunctional cobinamide kinase-cobinamide phosphate guanylyltransferase
J. Bacteriol.
173
6074-6087
1991
Pseudomonas denitrificans (nom. rej.) (P29930), Pseudomonas denitrificans (nom. rej.)
brenda
Sato, K.; Nakashima, T.; Shimizu, S.
Assay, purification and characterization of cob(I)alamin adenosyltransferase of Protaminobacter ruber
J. Nutr. Sci. Vitaminol.
30
405-413
1984
Serratia plymuthica
brenda
Suh, S.J.; Escalante-Semerena, J.C.
Purification and initial characterization of the ATP:corrinoid adenosyltransferase encoded by the cobA gene of Salmonella typhimurium
J. Bacteriol.
177
921-925
1995
Salmonella enterica subsp. enterica serovar Typhimurium
brenda
Fonseca, M.V.; Escalante-Semerena, J.C.
An in vitro reducing system for the enzymic conversion of cobalamin to adenosylcobalamin
J. Biol. Chem.
276
32101-32108
2001
Salmonella enterica
brenda
Johnson, C.L.; Pechonick, E.; Park, S.D.; Havemann, G.D.; Leal, N.A.; Bobik, T.A.
Functional genomic, biochemical, and genetic characterization of the Salmonella pduO gene, an ATP:cob(I)alamin adenosyltransferase gene
J. Bacteriol.
183
1577-1584
2001
Salmonella enterica, Salmonella enterica subsp. enterica serovar Typhimurium (P31570)
brenda
Fonseca, M.V.; Buan, N.R.; Horswill, A.R.; Rayment, I.; Escalante-Semerena, J.C.
The ATP:co(I)rrinoid adenosyltransferase (CobA) enzyme of Salmonella enterica requires the 2'-OH group of ATP for function and yields inorganic triphosphate as its reaction byproduct
J. Biol. Chem.
277
33127-33131
2002
Salmonella enterica
brenda
Leal, N.A.; Park, S.D.; Kima, P.E.; Bobik, T.A.
Identification of the human and bovine ATP:Cob(I)alamin adenosyltransferase cDNAs based on complementation of a bacterial mutant
J. Biol. Chem.
278
9227-9234
2003
Bos taurus, Homo sapiens (Q96EY8), Homo sapiens
brenda
Saridakis, V.; Yakunin, A.F.; Xu, X.; Anandakumar, P.; Pennycooke, M.; Gu, J.; Cheung, F.; Lew, J.M.; Sanishvili, N.; Joachimiak, A.; Arrowsmith, C.H.; Edwards, A.M.; Christendat, D.
The structural basis for methylmalonic aciduria. The crystal structure of archaeal ATP:cobalamin adenosyltransferase
J. Biol. Chem.
279
23646-23653
2004
Thermoplasma acidophilum (Q9HIA7), Thermoplasma acidophilum
brenda
Stich, T.A.; Yamanishi, M.; Banerjee, R.; Brunold, T.C.
Spectroscopic evidence for the formation of a four-coordinate Co2+ cobalamin species upon binding to the human ATP:cobalamin adenosyltransferase
J. Am. Chem. Soc.
127
7660-7661
2005
Homo sapiens
brenda
Stich, T.A.; Buan, N.R.; Escalante-Semerena, J.C.; Brunold, T.C.
Spectroscopic and computational studies of the ATP:corrinoid adenosyltransferase (CobA) from Salmonella enterica: insights into the mechanism of adenosylcobalamin biosynthesis
J. Am. Chem. Soc.
127
8710-8719
2005
Salmonella enterica
brenda
Buan, N.R.; Suh, S.J.; Escalante-Semerena, J.C.
The eutT gene of Salmonella enterica encodes an oxygen-labile, metal-containing ATP:corrinoid adenosyltransferase enzyme
J. Bacteriol.
186
5708-5714
2004
Salmonella enterica, Salmonella enterica subsp. enterica serovar Typhimurium (P31570)
brenda
Johnson, C.L.; Buszko, M.L.; Bobik, T.A.
Purification and initial characterization of the Salmonella enterica PduO ATP:Cob(I)alamin adenosyltransferase
J. Bacteriol.
186
7881-7887
2004
Salmonella enterica
brenda
Leal, N.A.; Olteanu, H.; Banerjee, R.; Bobik, T.A.
Human ATP:cob(I)alamin adenosyltransferase and its interaction with methionine synthase reductase
J. Biol. Chem.
279
47536-47542
2004
Homo sapiens
brenda
Schubert, H.L.; Hill, C.P.
Structure of ATP-bound human ATP:cobalamin adenosyltransferase
Biochemistry
45
15188-15196
2006
Homo sapiens (Q96EY8), Homo sapiens
brenda
Buan, N.R.; Rehfeld, K.; Escalante-Semerena, J.C.
Studies of the CobA-type ATP:Co(I)rrinoid adenosyltransferase enzyme of Methanosarcina mazei strain Go1
J. Bacteriol.
188
3543-3550
2006
Methanosarcina mazei
brenda
Buan, N.R.; Escalante-Semerena, J.C.
Computer-assisted docking of flavodoxin with the ATP:Co(I)rrinoid adenosyltransferase (CobA) enzyme reveals residues critical for protein-protein interactions but not for catalysis
J. Biol. Chem.
280
40948-40956
2005
Salmonella enterica
brenda
Buan, N.R.; Escalante-Semerena, J.C.
Purification and initial biochemical characterization of ATP:Cob(I)alamin adenosyltransferase (EutT) enzyme of Salmonella enterica
J. Biol. Chem.
281
16971-16977
2006
Salmonella enterica
brenda
St.Maurice, M.; Mera, P.E.; Taranto, M.P.; Sesma, F.; Escalante-Semerena, J.C.; Rayment, I.
Structural characterization of the active site of the PduO-type ATP:Co(I)rrinoid adenosyltransferase from Lactobacillus reuteri
J. Biol. Chem.
282
2596-2605
2007
Limosilactobacillus reuteri, Limosilactobacillus reuteri CRL1098
brenda
Erger, K.E.; Conlon, T.J.; Leal, N.A.; Zori, R.; Bobik, T.A.; Flotte, T.R.
In vivo expression of human ATP:cob(I)alamin adenosyltransferase (ATR) using recombinant adeno-associated virus (rAAV) serotypes 2 and 8
J. Gene Med.
9
462-469
2007
Homo sapiens
brenda
Zhang, J.; Dobson, C.M.; Wu, X.; Lerner-Ellis, J.; Rosenblatt, D.S.; Gravel, R.A.
Impact of cblB mutations on the function of ATP:cob(I)alamin adenosyltransferase in disorders of vitamin B12 metabolism
Mol. Genet. Metab.
87
315-322
2006
Homo sapiens
brenda
Tanaka, Y.; Sasaki, T.; Kumagai, I.; Yasutake, Y.; Yao, M.; Tanaka, I.; Tsumoto, K.
Molecular properties of two proteins homologous to PduO-type ATP:cob(I)alamin adenosyltransferase from Sulfolobus tokodaii.
Proteins
68
446-457
2007
Sulfurisphaera tokodaii, Sulfurisphaera tokodaii (Q970Z7), Sulfurisphaera tokodaii 7 (Q970Z7)
brenda
Park, A.K.; Moon, J.H.; Lee, S.H.; Chi, Y.M.
Crystallization and preliminary X-ray crystallographic studies of a PduO-type ATP:cob(I)alamin adenosyltransferase from Bacillus cereus
Acta Crystallogr. Sect. F
64
648-650
2008
Bacillus cereus
brenda
Mera, P.E.; St Maurice, M.; Rayment, I.; Escalante-Semerena, J.C.
Structural and functional analyses of the human-type corrinoid adenosyltransferase (PduO) from Lactobacillus reuteri
Biochemistry
46
13829-13836
2007
Limosilactobacillus reuteri
brenda
Fan, C.; Bobik, T.A.
Functional characterization and mutation analysis of human ATP:Cob(I)alamin adenosyltransferase
Biochemistry
47
2806-2813
2008
Homo sapiens (Q96EY8), Homo sapiens
brenda
St Maurice, M.; Mera, P.; Park, K.; Brunold, T.C.; Escalante-Semerena, J.C.; Rayment, I.
Structural characterization of a human-type corrinoid adenosyltransferase confirms that coenzyme B12 is synthesized through a four-coordinate intermediate
Biochemistry
47
5755-5766
2008
Limosilactobacillus reuteri
brenda
Moon, J.H.; Park, A.K.; Jang, E.H.; Kim, H.S.; Chi, Y.M.
Crystal structure of a PduO-type ATP: cobalamin adenosyltransferase from Burkholderia thailandensis
Proteins
72
1066-1070
2008
Burkholderia thailandensis (Q2SZ09), Burkholderia thailandensis
brenda
Medina, C.; Crespo-Rivas, J.; Moreno, J.; Espuny, M.; Cubo, M.
Mutation in the cobO gene generates auxotrophy for cobalamin and methionine and impairs the symbiotic properties of Sinorhizobium fredii HH103 with soybean and other legumes
Arch. Microbiol.
191
11-21
2009
Sinorhizobium fredii
brenda
Park, K.; Mera, P.; Escalante-Semerena, J.; Brunold, T.
Kinetic and spectroscopic studies of the ATP:corrinoid adenosyltransferase PduO from Lactobacillus reuteri: Substrate specificity and insights into the mechanism of Co(II)corrinoid reduction
Biochemistry
47
9007-9015
2008
Limosilactobacillus reuteri
brenda
Mera, P.; Maurice, M.; Rayment, I.; Escalante-Semerena, J.
Residue Phe112 of the human-type corrinoid adenosyltransferase (PduO) enzyme of lactobacillus reuteri is critical to the formation of the four-coordinate Co(II) corrinoid substrate and to the activity of the enzyme
Biochemistry
48
3138-3145
2009
Limosilactobacillus reuteri
brenda
Padovani, D.; Banerjee, R.
A rotary mechanism for coenzyme B(12) synthesis by adenosyltransferase
Biochemistry
48
5350-5357
2009
Methylorubrum extorquens
brenda
Mera, P.; Escalante-Semerena, J.
Dihydroflavin-driven adenosylation of 4-coordinate Co(II) corrinoids: are cobalamin reductases enzymes or electron transfer proteins?
J. Biol. Chem.
285
2911-2917
2010
Homo sapiens, Limosilactobacillus reuteri
brenda
Zhang, J.; Wu, X.; Padovani, D.; Schubert, H.; Gravel, R.
Ligand-binding by catalytically inactive mutants of the cblB complementation group defective in human ATP:cob(I)alamin adenosyltransferase
Mol. Genet. Metab.
98
278-284
2009
Homo sapiens
brenda
Park, A.K.; Chi, Y.M.; Moon, J.H.
Crystal structure of PduO-Type ATP:Cob(I)alamin adenosyltransferase from Bacillus cereus in a complex with ATP
Biochem. Biophys. Res. Commun.
408
417-421
2011
Bacillus cereus
brenda
Moore, T.C.; Newmister, S.A.; Rayment, I.; Escalante-Semerena, J.C.
Structural insights into the mechanism of four-coordinate Cob(II)alamin formation in the active site of the Salmonella enterica ATP:Co(I)rrinoid adenosyltransferase enzyme: critical role of residues Phe91 and Trp93
Biochemistry
51
9647-9657
2012
Salmonella enterica
brenda
Park, K.; Mera, P.E.; Escalante-Semerena, J.C.; Brunold, T.C.
Spectroscopic characterization of active-site variants of the PduO-type ATP:corrinoid adenosyltransferase from Lactobacillus reuteri: insights into the mechanism of four-coordinate Co(II)corrinoid formation
Inorg. Chem.
51
4482-4494
2012
Limosilactobacillus reuteri
brenda
Park, K.; Mera, P.E.; Moore, T.C.; Escalante-Semerena, J.C.; Brunold, T.C.
Unprecedented mechanism employed by the Salmonella enterica EutT ATP:Co(I)rrinoid adenosyltransferase precludes adenosylation of incomplete Co(II)rrinoids
Angew. Chem. Int. Ed. Engl.
54
7158-7161
2015
Salmonella enterica
brenda
Pallares, I.G.; Moore, T.C.; Escalante-Semerena, J.C.; Brunold, T.C.
Spectroscopic studies of the Salmonella enterica adenosyltransferase enzyme SeCobA: molecular-level insight into the mechanism of substrate Cob(II)alamin activation
Biochemistry
53
7969-7982
2014
Salmonella enterica (P31570), Salmonella enterica
brenda
Pallares, I.G.; Moore, T.C.; Escalante-Semerena, J.C.; Brunold, T.C.
Spectroscopic Studies of the EutT Adenosyltransferase from Salmonella enterica: Mechanism of Four-Coordinate Co(II)Cbl Formation
J. Am. Chem. Soc.
138
3694-3704
2016
Salmonella enterica
brenda
Moore, T.C.; Mera, P.E.; Escalante-Semerena, J.C.
The EutT enzyme of Salmonella enterica is a unique ATP:Cob(I)alamin adenosyltransferase metalloprotein that requires ferrous ions for maximal activity
J. Bacteriol.
196
903-910
2014
Salmonella enterica, Salmonella enterica JE6583
brenda
Costa, F.G.; Escalante-Semerena, J.C.
A new class of EutT ATP Co(I)rrinoid adenosyltransferases found in Listeria monocytogenes and other Firmicutes does not require a metal ion for activity
Biochemistry
57
5076-5087
2018
Listeria monocytogenes (A0A0H3GFM5), Listeria monocytogenes 10403S (A0A0H3GFM5)
brenda
Stracey, N.G.; Costa, F.G.; Escalante-Semerena, J.C.; Brunold, T.C.
Spectroscopic study of the EutT adenosyltransferase from Listeria monocytogenes evidence for the formation of a four-coordinate cob(II)alamin intermediate
Biochemistry
57
5088-5095
2018
Listeria monocytogenes
brenda
Costa, F.G.; Greenhalgh, E.D.; Brunold, T.C.; Escalante-Semerena, J.C.
Mutational and functional analyses of substrate binding and catalysis of the Listeria monocytogenes EutT ATP Co(I)rrinoid adenosyltransferase
Biochemistry
59
1124-1136
2020
Listeria monocytogenes (A0A0E1R5H0), Listeria monocytogenes, Listeria monocytogenes LL195 (A0A0E1R5H0)
brenda
Ortiz de Orue Lucana, D.; Hickey, N.; Hensel, M.; Klare, J.P.; Geremia, S.; Tiufiakova, T.; Torda, A.E.
The crystal structure of the C-terminal domain of the Salmonella enterica PduO protein an old fold with a new heme-binding mode
Front. Microbiol.
7
1010
2016
Salmonella enterica (A0A5Z8GIG6), Salmonella enterica
brenda
Park, K.; Mera, P.E.; Escalante-Semerena, J.C.; Brunold, T.C.
Resonance Raman spectroscopic study of the interaction between Co(II)rrinoids and the ATP corrinoid adenosyltransferase PduO from Lactobacillus reuteri
J. Biol. Inorg. Chem.
21
669-681
2016
Limosilactobacillus reuteri (Q50EJ2), Limosilactobacillus reuteri
brenda