Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.21 extracted from

  • Bohren, K.M.; Brownlee, J.M.; Milne, A.C.; Gabbay, K.H.; Harrison, D.H.
    The structure of Apo R268A human aldose reductase: Hinges and latches that control the kinetic mechanism (2005), Biochim. Biophys. Acta, 1748, 201-212.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21 Homo sapiens

Crystallization (Commentary)

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

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 [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/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
aldose + NADPH + H+ Homo sapiens
-
alditol + NADP+
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 Homo sapiens

Reaction

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 and Products (Substrate)

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

Subunits Comment Organism
More the cofactor binding loop structure influences the kinetic mechanism of the enzyme, loop movement, overview Homo sapiens

Synonyms

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Homo sapiens

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Homo sapiens

Cofactor

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