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Literature summary for 1.8.1.4 extracted from

  • Liu, T.C.; Soo Hong, Y.; Korotchkina, L.G.; Vettakkorumakankav, N.N.; Patel, M.S.
    Site-directed mutagenesis of human dihydrolipoamide dehydrogenase: role of lysine-54 and glutamate-192 in stabilizing the thiolate-FAD intermediate (1999), Protein Expr. Purif., 16, 27-39.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
wild-type and mutant enzymes K54E, S53K54-K53S54 and E192Q, overexpression in Escherichia coli Homo sapiens

Protein Variants

Protein Variants Comment Organism
E192Q specific activity is markedly decreased, less than 5% of the wild-type activity, Km-values for lipoamide and dihydrolipoamide are markedly reduced Homo sapiens
K54E about 25% less bound FAD compared to wild-type, specific activity is markedly decreased, less than 5% of the wild-type activity, Km-value for lipoamide is increased by about twofold Homo sapiens
S53K/K54S about 25% less bound FAD compared to wild-type, specific activity is markedly decreased, less than 5% of the wild-type activity, Km-values for lipoamide and dihydrolipoamide are markedly reduced. The catalytic rate constant, turnover number/Km, is significantly lower than wild-type Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05
-
dihydrolipoamide mutant enzyme S53K/K54S Homo sapiens
0.071
-
NADH mutant enzyme E192Q Homo sapiens
0.12
-
dihydrolipoamide mutant enzyme E192Q Homo sapiens
0.13
-
NADH mutant enzyme S53K/K54S Homo sapiens
0.31
-
NAD+ mutant enzyme E192Q Homo sapiens
0.31
-
Lipoamide mutant enzyme E192Q Homo sapiens
0.4
-
NAD+ wild-type enzyme Homo sapiens
0.55
-
NAD+ mutant enzyme S53K/K54S Homo sapiens
0.63
-
Lipoamide mutant enzyme S53K/K54S Homo sapiens
0.88
-
dihydrolipoamide wild-type enzyme Homo sapiens
2.25
-
Lipoamide wild-type enzyme Homo sapiens
5
-
Lipoamide mutant enzyme K54E Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutant enzymes K54E, S53K54-K53S54 and E192Q Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dihydrolipoamide + NAD+
-
Homo sapiens lipoamide + NADH
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.75
-
NAD+ mutant enzyme S53K/K54S Homo sapiens
8.5
-
dihydrolipoamide mutant enzyme S53K/K54S Homo sapiens
14.4
-
Lipoamide mutant enzyme S53K/K54S Homo sapiens
19.8
-
NADH mutant enzyme S53K/K54S Homo sapiens
23
-
Lipoamide mutant enzyme K54E Homo sapiens
26.2
-
NADH mutant enzyme E192Q Homo sapiens
29.7
-
dihydrolipoamide mutant enzyme E192Q Homo sapiens
30
-
NAD+ mutant enzyme E192Q Homo sapiens
81.3
-
Lipoamide mutant enzyme E192Q Homo sapiens
340
-
NADH wild-type enzyme Homo sapiens
567
-
dihydrolipoamide wild-type enzyme Homo sapiens
574
-
NAD+ wild-type enzyme Homo sapiens
649
-
Lipoamide wild-type enzyme Homo sapiens

Cofactor

Cofactor Comment Organism Structure
FAD wild-type enzyme contains 1 FAD per subunit, mutant enzymes K54E and S53K/K54S have about 25% less bound FAD Homo sapiens
NAD+
-
Homo sapiens