Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
in mutant E161Q, the hydrolysis step becomes completely rate-limiting so that a thioester enzyme intermediate accumulates at steady state. Crystallization of mutant E161Q in the presence of 5 mM UDP-glucose and 2 mM NAD results in trapping a thiohemiacetal enzyme intermediate. Residue Cys276 is covalently modified in the structure, establishing its role as catalytic nucleophile of the reaction | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
D280A | extremely poor enzymic activity | Homo sapiens |
E161Q | hydrolysis step becomes completely rate-limiting so that a thioester enzyme intermediate accumulates at steady state. Crystallization of E161Q in the presence of 5 mM UDP-glucose and 2 mM NAD results in trapping a thiohemiacetal enzyme intermediate | Homo sapiens |
K220A | extremely poor enzymic activity | Homo sapiens |
N224A | steady-state kinetic parameters are within an order of magnitude of the native enzyme | Homo sapiens |
T131S | steady-state kinetic parameters are within an order of magnitude of the native enzyme | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.035 | - |
UDP-alpha-D-glucose | wild-type, pH 8.7, 25°C | Homo sapiens | |
0.055 | - |
UDP-alpha-D-glucose | mutant E161Q, pH 8.7, 25°C | Homo sapiens | |
0.1 | - |
NAD+ | mutant E161Q, pH 8.7, 25°C | Homo sapiens | |
0.7 | - |
NAD+ | wild-type, pH 8.7, 25°C | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | O60701 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-alpha-D-glucose + 2 NAD+ + H2O | - |
Homo sapiens | UDP-alpha-D-glucuronate + 2 NADH + 2 H+ | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0014 | - |
UDP-alpha-D-glucose | mutant E161Q, pH 8.7, 25°C | Homo sapiens | |
0.85 | - |
UDP-alpha-D-glucose | wild-type, pH 8.7, 25°C | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.014 | - |
NAD+ | mutant E161Q, pH 8.7, 25°C | Homo sapiens | |
0.025 | - |
UDP-alpha-D-glucose | mutant E161Q, pH 8.7, 25°C | Homo sapiens | |
1.2 | - |
NAD+ | wild-type, pH 8.7, 25°C | Homo sapiens | |
24 | - |
UDP-alpha-D-glucose | wild-type, pH 8.7, 25°C | Homo sapiens |