Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Escherichia coli |
gene yihU, phylogenetic analysis and tree | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
apoenzyme, hanging drop vapor diffusion method, using 25% (w/v) PEG (polyethylene glycol) 1500, 0.1 M sodium malonate dibasic monohydrate, imidazole, boric acid buffer pH 4.0. NADH-bound enzyme, hanging drop vapor diffusion method, using 25% (w/v) PEG (polyethylene glycol) 3350, 0.2 M CH3COONH4, 0.1 M Bis-Tris pH 6.5. NADH- and (S)-2,3-dihydroxypropane-1-sulfonate-bound enzyme, hanging drop vapor diffusion method, using 32% (w/v) PEG 3350, 0.2 mM CH3COONH4, 0.1 M Bis-Tris pH 6.5 | Escherichia coli |
purified enzyme in its apo and cofactor-bound states, as well as in ternary complex YihU-NADH-DHPS with the cofactor and product bound in the active site, X-ray diffraction structure determination and analysis | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
G122S | the mutant shows reduced activity compared to the wild type enzyme | Escherichia coli |
G122S | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme | Escherichia coli |
R123G | the mutant shows reduced activity compared to the wild type enzyme | Escherichia coli |
R123G | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme | Escherichia coli |
T124G | the mutant shows reduced activity compared to the wild type enzyme | Escherichia coli |
T124G | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
hexahydro-NADH | - |
Escherichia coli | |
hexahydronicotinamide adenine dinucleotide | - |
Escherichia coli | |
additional information | modified NADH analogues are inhibitors of YihU | Escherichia coli | |
tetrahydro-NADH | - |
Escherichia coli | |
tetrahydronicotinamide adenine dinucleotide | - |
Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics analysis and kinetic mechanism, overview | Escherichia coli | |
0.009 | - |
NADH | enzyme mutant R123G, pH and temperature not specified in the publication | Escherichia coli | |
0.009 | - |
NADH | mutant enzyme R123G, at pH 8.0 and 30°C | Escherichia coli | |
0.023 | - |
NADH | enzyme mutant T124G, pH and temperature not specified in the publication | Escherichia coli | |
0.023 | - |
NADH | mutant enzyme T124G, at pH 8.0 and 30°C | Escherichia coli | |
0.082 | - |
NADH | wild type enzyme, at pH 8.0 and 30°C | Escherichia coli | |
0.082 | - |
NADH | wild-type enzyme, pH and temperature not specified in the publication | Escherichia coli | |
0.123 | - |
NADH | enzyme mutant G122S, pH and temperature not specified in the publication | Escherichia coli | |
0.123 | - |
NADH | mutant enzyme G122S, at pH 8.0 and 30°C | Escherichia coli | |
0.3 | - |
(S)-3-sulfolactaldehyde | wild type enzyme, at pH 8.0 and 30°C | Escherichia coli | |
0.3 | - |
2-hydroxy-3-oxopropane-1-sulfonate | wild-type enzyme, pH and temperature not specified in the publication | Escherichia coli | |
1.63 | - |
2-hydroxy-3-oxopropane-1-sulfonate | enzyme mutant G122S, pH and temperature not specified in the publication | Escherichia coli | |
1.63 | - |
(S)-3-sulfolactaldehyde | mutant enzyme G122S, at pH 8.0 and 30°C | Escherichia coli | |
5.74 | - |
2-hydroxy-3-oxopropane-1-sulfonate | enzyme mutant T124G, pH and temperature not specified in the publication | Escherichia coli | |
5.74 | - |
(S)-3-sulfolactaldehyde | mutant enzyme T124G, at pH 8.0 and 30°C | Escherichia coli | |
7.99 | - |
2-hydroxy-3-oxopropane-1-sulfonate | enzyme mutant R123G, pH and temperature not specified in the publication | Escherichia coli | |
7.99 | - |
(S)-3-sulfolactaldehyde | mutant enzyme R123G, at pH 8.0 and 30°C | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
123000 | - |
gel filtration | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-3-sulfolactaldehyde + NADH + H+ | Escherichia coli | - |
(S)-2,3-dihydroxypropane-1-sulfonate + NAD+ | - |
? | |
2-hydroxy-3-oxopropane-1-sulfonate + NADH + H+ | Escherichia coli | - |
2,3-dihydroxypropane-1-sulfonate + NAD+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A9V8 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography and Superdex 75 gel filtration | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2,3-dihydroxypropane-1-sulfonate + NAD+ = 2-hydroxy-3-oxopropane-1-sulfonate + NADH + H+ | rapid equilibrium sequential Bi-Bi mechanism, dynamic domain movements occurs during catalysis. A conserved sulfonate pocket in SLA reductase recognizes the sulfonate oxygens through hydrogen bonding to Asn174, Ser178, and the backbone amide of Arg123, along with an ordered water molecule | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-3-sulfolactaldehyde + NADH + H+ | - |
Escherichia coli | (S)-2,3-dihydroxypropane-1-sulfonate + NAD+ | - |
? | |
2-hydroxy-3-oxopropane-1-sulfonate + NADH + H+ | - |
Escherichia coli | 2,3-dihydroxypropane-1-sulfonate + NAD+ | - |
r | |
additional information | the SLA reductase shows no detectable activity on the analogous glycolytic intermediate glyceraldehyde-3-phosphate. Substrate binding analysis | Escherichia coli | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
homotetramer | 4 * 30000, calculated from amino acid sequence | Escherichia coli |
tetramer | YihU forms a dimer of intimate homodimer pairs. The enzyme exuísts as a tetramer in crystal and in solution. Within the asymmetric unit, each protomer adopts a two-domain architecture containing an N-terminal nucleotide-binding domain (residues 1-164) and a C-terminal helical bundle (residues 165-294) both connected by the long interdomain helix alpha8. The N-terminal domain is composed of a classical alpha/beta Rossmann fold (composed of an extended sheet formed by beta1-6, flanked by alpha1-5 in a three-layered sandwich) appended with an additional beta-alpha-beta motif containing beta7-9 and alpha6. Four interfaces are present within the subunits of the YihU tetramer. Detailed structure analysis, overview | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
3-sulfolactaldehyde reductase | UniProt | Escherichia coli |
NADH-dependent SLA reductase | - |
Escherichia coli |
SLA reductase | - |
Escherichia coli |
sulfolactaldehyde reductase | - |
Escherichia coli |
yihU | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
12.7 | - |
NADH | enzyme mutant T124G, pH and temperature not specified in the publication | Escherichia coli | |
12.7 | - |
NADH | mutant enzyme T124G, at pH 8.0 and 30°C | Escherichia coli | |
22.1 | - |
NADH | enzyme mutant R123G, pH and temperature not specified in the publication | Escherichia coli | |
22.1 | - |
NADH | mutant enzyme R123G, at pH 8.0 and 30°C | Escherichia coli | |
27 | - |
2-hydroxy-3-oxopropane-1-sulfonate | enzyme mutant T124G, pH and temperature not specified in the publication | Escherichia coli | |
27 | - |
(S)-3-sulfolactaldehyde | mutant enzyme T124G, at pH 8.0 and 30°C | Escherichia coli | |
68 | - |
2-hydroxy-3-oxopropane-1-sulfonate | enzyme mutant R123G, pH and temperature not specified in the publication | Escherichia coli | |
68 | - |
(S)-3-sulfolactaldehyde | mutant enzyme R123G, at pH 8.0 and 30°C | Escherichia coli | |
71 | - |
2-hydroxy-3-oxopropane-1-sulfonate | enzyme mutant G122S, pH and temperature not specified in the publication | Escherichia coli | |
71 | - |
(S)-3-sulfolactaldehyde | mutant enzyme G122S, at pH 8.0 and 30°C | Escherichia coli | |
178 | - |
NADH | enzyme mutant G122S, pH and temperature not specified in the publication | Escherichia coli | |
178 | - |
NADH | mutant enzyme G122S, at pH 8.0 and 30°C | Escherichia coli | |
332 | - |
2-hydroxy-3-oxopropane-1-sulfonate | wild-type enzyme, pH and temperature not specified in the publication | Escherichia coli | |
332 | - |
(S)-3-sulfolactaldehyde | wild type enzyme, at pH 8.0 and 30°C | Escherichia coli | |
548 | - |
NADH | wild-type enzyme, pH and temperature not specified in the publication | Escherichia coli | |
548 | - |
NADH | wild type enzyme, at pH 8.0 and 30°C | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | cofactor binding analysis | Escherichia coli | |
NAD+ | - |
Escherichia coli | |
NADH | - |
Escherichia coli | |
NADH | dependent on | Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
1.81 | - |
tetrahydronicotinamide adenine dinucleotide | wild type enzyme, at pH 8.0 and 30°C | Escherichia coli | |
4.63 | - |
hexahydronicotinamide adenine dinucleotide | wild type enzyme, at pH 8.0 and 30°C | Escherichia coli |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
4.03 | - |
wild-type enzyme, pH and temperature not specified in the publication | Escherichia coli | tetrahydro-NADH | |
4.03 | - |
wild type enzyme, at pH 8.0 and 30°C | Escherichia coli | tetrahydronicotinamide adenine dinucleotide | |
10.3 | - |
wild-type enzyme, pH and temperature not specified in the publication | Escherichia coli | hexahydro-NADH | |
10.3 | - |
wild type enzyme, at pH 8.0 and 30°C | Escherichia coli | hexahydronicotinamide adenine dinucleotide |
General Information | Comment | Organism |
---|---|---|
evolution | based on conserved sequence motifs, enzyme SLA reductase is assigned to the beta-hydroxyacid dehydrogenase (beta-HAD) family, comparisons of typical sequence motifs. SLA reductases form a separate subgroup with conserved sulfonate substrate-binding sequence at residues 122-124 (YihU numbering). SLA reductases share beta-HAD sequence motifs but possess a unique sulfonate substrate-binding sequence (extended motif-2 [D/EVPVGRTXX-XAXXG]) | Escherichia coli |
additional information | a conserved sulfonate pocket in SLA reductase recognizes the sulfonate oxygens through hydrogen bonding to Asn174, Ser178, and the backbone amide of Arg123, along with an ordered water molecule. This triad of residues distinguishes these enzymes from classical beta-HADs that act on carboxylate substrates. Determination of the structural basis for cofactor binding and sulfonate recognition, dynamic structural changes occur during catalysis | Escherichia coli |
physiological function | 2,3-dihydroxypropanesulfonate (DHPS) is a major sulfur species in the biosphere. One important route for the production of DHPS is sulfoglycolytic catabolism of sulfoquinovose (SQ) through the Embden-Meyerhof-Parnas (sulfo-EMP) pathway. SQ is a sulfonated carbohydrate present in plant and cyanobacterial sulfolipids (sulfoquinovosyl diacylglyceride and its metabolites) and is biosynthesized globally at a rate of around 10 billion tons per annum. The final step in the bacterial sulfo-EMP pathway involves reduction of sulfolactaldehyde (SLA) to DHPS, catalyzed by the NADH-dependent SLA reductase | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4.7 | - |
2-hydroxy-3-oxopropane-1-sulfonate | enzyme mutant T124G, pH and temperature not specified in the publication | Escherichia coli | |
4.7 | - |
(S)-3-sulfolactaldehyde | mutant enzyme T124G, at pH 8.0 and 30°C | Escherichia coli | |
8.5 | - |
2-hydroxy-3-oxopropane-1-sulfonate | enzyme mutant R123G, pH and temperature not specified in the publication | Escherichia coli | |
8.5 | - |
(S)-3-sulfolactaldehyde | mutant enzyme R123G, at pH 8.0 and 30°C | Escherichia coli | |
43 | - |
(S)-3-sulfolactaldehyde | mutant enzyme G122S, at pH 8.0 and 30°C | Escherichia coli | |
43.6 | - |
2-hydroxy-3-oxopropane-1-sulfonate | enzyme mutant G122S, pH and temperature not specified in the publication | Escherichia coli | |
552.2 | - |
NADH | enzyme mutant T124G, pH and temperature not specified in the publication | Escherichia coli | |
570 | - |
NADH | mutant enzyme T124G, at pH 8.0 and 30°C | Escherichia coli | |
1090 | - |
(S)-3-sulfolactaldehyde | wild type enzyme, at pH 8.0 and 30°C | Escherichia coli | |
1106.7 | - |
2-hydroxy-3-oxopropane-1-sulfonate | wild-type enzyme, pH and temperature not specified in the publication | Escherichia coli | |
1447.2 | - |
NADH | enzyme mutant G122S, pH and temperature not specified in the publication | Escherichia coli | |
1450 | - |
NADH | mutant enzyme G122S, at pH 8.0 and 30°C | Escherichia coli | |
2360 | - |
NADH | mutant enzyme R123G, at pH 8.0 and 30°C | Escherichia coli | |
2455.6 | - |
NADH | enzyme mutant R123G, pH and temperature not specified in the publication | Escherichia coli | |
6682.9 | - |
NADH | wild-type enzyme, pH and temperature not specified in the publication | Escherichia coli | |
6720 | - |
NADH | wild type enzyme, at pH 8.0 and 30°C | Escherichia coli |