Cloned (Comment) | Organism |
---|---|
expression of histidine and cysteine mutant alpha subunits together with the wild-type beta subunit in insect cells by means of baculovirus vectors | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
C150S | the mutation has no major effect on tetramer assembly, but the amount of tetramer is slightly reduced, being about 80% of that of the wild-type enzyme | Homo sapiens |
C486S | the mutation totally prevents tetramer assembly | Homo sapiens |
C511S | the mutation totally prevents tetramer assembly | Homo sapiens |
H141S | the mutation has no effect on enzyme activity and does not inhibit tetramer assembly | Homo sapiens |
H165S | the mutation produces a reduction of about 60% in enzyme activity per unit extractable cell protein relative to that obtained with the wild-type alpha subunit, the amount of tetramer is reduced by about 20-25%, the Km values for Fe2+, 2-oxoglutarate, ascorbate and the peptide substrate with the mutant are identical to those with the wild-type enzyme | Homo sapiens |
H221S | the mutation produces a reduction of about 30% in enzyme activity per unit extractable cell protein relative to that obtained with the wild-type alpha subunit, the amount of tetramer is reduced by about 20-25%, the Km values for Fe2+, 2-oxoglutarate, ascorbate and the peptide substrate with the mutant are identical to those with the wild-type enzyme | Homo sapiens |
H296S | the mutation has no effect on enzyme activity and does not inhibit tetramer assembly | Homo sapiens |
H324S | the mutation totally prevents tetramer assembly | Homo sapiens |
H412S | the mutation causes a complete inactivation of the enzyme with no effect on tetramer assembly or binding of the tetramer to poly(L-proline), role in the binding of Fe2+ to a catalytic site | Homo sapiens |
H483S | the mutation causes a complete inactivation of the enzyme with no effect on tetramer assembly or binding of the tetramer to poly(L-proline), role in the binding of Fe2+ to a catalytic site | Homo sapiens |
H501S | the mutation reduces the enzyme activity to about 4% with no effect on tetramer assembly or binding of the tetramer to poly(L-proline), role in the binding of Fe2+ to a catalytic site, the Km values for Fe2+, ascorbate and the peptide substrate with the mutant are identical to those with the wild-type enzyme, but the Km for 2-oxoglutarate is about 2.5fold higher. The main difference is that the Vmax determined from kinetic plots is consistently less than about 5% of that of the wild-type enzyme | Homo sapiens |
H63S | the mutation has no effect on enzyme activity and does not inhibit tetramer assembly | Homo sapiens |
N96Q/N242Q | the amount of enzyme activity observed with the double mutant alpha subunit is identical to that of the wild-type enzyme, the size of the double mutant alpha subunit is distinctly smaller than that of either the diglycosylated or monoglycosylated alpha subunit present in the wild-type enzyme, the difference being consistent with loss of all the carbohydrate | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Pyridine 2,4-dicarboxylate | inhibits wild-type enzyme and enzyme tetramer containing the histidine 501 to serine mutant alpha subunit | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.018 | - |
(Pro-Pro-Gly)10 | wild-type enzyme tetramer and enzyme tetramer containing the H501S mutant alpha subunit | Homo sapiens | |
0.05 | - |
2-oxoglutarate | wild-type enzyme tetramer | Homo sapiens | |
0.15 | - |
2-oxoglutarate | enzyme tetramer containing the H501S mutant alpha subunit | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | enzyme tetramer containing the histidine 501 to serine mutant alpha subunit: Km 0.005 mM | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
enzyme activity of Triton X-100 extracts from cells expressing various mutant alpha subunits together with the wild-type protein disulfide-isomerase/beta subunit | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(Pro-Pro-Gly)10 + 2-oxoglutarate + O2 | - |
Homo sapiens | ? + succinate + CO2 | - |
? | |
(Pro-Pro-Gly)n + 2-oxoglutarate + O2 | n: 1,5,10 | Homo sapiens | (Pro-4-hydroxy-Pro-Gly)n + succinate + CO2 | n: 1,5,10 | ? |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ascorbate | Km for wild-type enzyme tetramer: 0.33 mM, enzyme tetramer containing the histidine 501 to serine mutant alpha subunit, Km: 0.4 mM | Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.002 | - |
Pyridine 2,4-dicarboxylate | wild-type enzyme tetramer | Homo sapiens | |
0.005 | - |
Pyridine 2,4-dicarboxylate | enzyme tetramer containing the histidine 501 to serine mutant alpha subunit | Homo sapiens |