Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain BL21 | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant R268A mutant enzyme, hanging drop vapour diffusion method, 30 mg/ml protein in 5 mM phosphate, pH 7.0, mixing with equal volume of well solution containing 20% v/v PEG 6000, 25 mM MES, pH 6.0, and 16 mM ammonium sulfate, 3 weeks, X-ray diffraction structure determination and analysis at 2.8 A resolution | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
C298A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
G213P | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
G213S | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
additional information | construction of deletion mutant DELTA214-219 showing reduced activity compared to the wild-type enzyme | Homo sapiens |
R268A | site-directed mutagenesis, the mutant shows similar kinetics for the aldehyde substrate, but highly reduced affinity for the nucleotide cofactor, and reduced activity compared to the wild-type enzyme | Homo sapiens |
R293A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
S210A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
S214A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
S226A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
W219A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
W219E | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant wild-type enzyme | Homo sapiens | |
0.02 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant R293A | Homo sapiens | |
0.02 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant W219A | Homo sapiens | |
0.02 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant W219E | Homo sapiens | |
0.03 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant S226A | Homo sapiens | |
0.05 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant R268A | Homo sapiens | |
0.11 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant S210A | Homo sapiens | |
0.13 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant S214A | Homo sapiens | |
0.42 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant G213S | Homo sapiens | |
0.5 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant DELTA214-219 | Homo sapiens | |
0.6 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant C298A | Homo sapiens | |
3.7 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant G213P | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
aldose + NADPH + H+ | Homo sapiens | - |
alditol + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 | Homo sapiens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
alditol + NAD(P)+ = aldose + NAD(P)H + H+ | the ordered bi-bi kinetic mechanism is controlled by hinges and latches of enzyme structure, residues R268, G213, and S226 are involved | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
aldose + NADPH + H+ | - |
Homo sapiens | alditol + NADP+ | - |
? | |
DL-glyceraldehyde + NADPH + H+ | - |
Homo sapiens | glycerol + NADP+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
More | the cofactor binding loop structure influences the kinetic mechanism of the enzyme, loop movement, overview | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
AKR1B1 | - |
Homo sapiens |
aldose reductase | - |
Homo sapiens |
More | the enzyme is a member of the aldo-keto reductase family | Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.09 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant W219E | Homo sapiens | |
0.16 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant R293A | Homo sapiens | |
0.32 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant W219A | Homo sapiens | |
0.38 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant R268A | Homo sapiens | |
0.5 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant wild-type enzyme | Homo sapiens | |
0.54 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant S226A | Homo sapiens | |
2 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant S210A | Homo sapiens | |
2.1 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant S214A | Homo sapiens | |
2.15 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant DELTA214-219 | Homo sapiens | |
3.7 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant G213S | Homo sapiens | |
4 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant C298A | Homo sapiens | |
4.6 | - |
DL-glyceraldehyde | pH 7.0, 25°C, recombinant mutant G213P | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | dependent on, several factors act to stabilize the NADPH-holding loop in either the open or closed conformation, residues R268, G213, and S226 are involved, loop movement, overview | Homo sapiens |