Protein Variants | Comment | Organism |
---|---|---|
E314A | site-directed mutagenesis | Homo sapiens |
E314A/S346I/K347D/K362H | site-directed mutagenesis, the quadruple mutant enzyme is a mainly NAD+-utilizing enzyme by a considerable increase in catalysis using NAD+ as the cofactor, shows increased inhibition by ATP compared to the wild-type enzyme | Homo sapiens |
E314A/S346K | site-directed mutagenesis | Homo sapiens |
E314A/S346K/K347Y/K362H | site-directed mutagenesis, the quadruple mutant enzyme is a mainly NAD+-utilizing enzyme by a considerable increase in catalysis using NAD+ as the cofactor, shows increased inhibition by ATP compared to the wild-type enzyme | Homo sapiens |
E314A/S346K/K347Y/K362Q | site-directed mutagenesis, the quadruple mutant enzyme is a mainly NAD+-utilizing enzyme by a considerable increase in catalysis using NAD+ as the cofactor, shows increased inhibition by ATP compared to the wild-type enzyme | Homo sapiens |
K347Y | site-directed mutagenesis, the mutant enzyme shows a 5fold increased Km for NADP+ compared to the wild-type enzyme | Homo sapiens |
K347Y/K362Q | site-directed mutagenesis | Homo sapiens |
K362H | site-directed mutagenesis | Homo sapiens |
K362Q | site-directed mutagenesis, the mutant enzyme displays a significant, over 140fold elevation in Km,NADP value compared with that of wild-type c-NADP-ME but no significant changes in the kcat,NADP value | Homo sapiens |
additional information | a series of E314A-containing c-NADP-ME quadruple mutants are changed to NAD+-utilizing enzymes by abrogating NADP+ binding and increasing NAD+ binding. Abolishing the repulsive effect of Glu314 in the quadruple mutants increases the binding affinity of NAD+ | Homo sapiens |
S346I/K347D/K362H | site-directed mutagenesis, the triple c-NADP-ME mutant does not show significant reduction in its Km,NAD values. This mutant exclusively utilizes NAD+ as its cofactor | Homo sapiens |
S346K | site-directed mutagenesis, site-directed mutagenesis, the mutant enzyme shows a 3fold increased Km for NADP+ compared to the wild-type enzyme | Homo sapiens |
S346K/K347Y | site-directed mutagenesis, the double mutant enzyme shows a 30fold increased Km for NADP+ compared to the wild-type enzyme | Homo sapiens |
S346K/K347Y/K362H | site-directed mutagenesis, the triple c-NADP-ME mutant does not show significant reduction in its Km,NAD values, but displays an enhanced value for kcat,NAD | Homo sapiens |
S346K/K347Y/K362Q | site-directed mutagenesis, the triple c-NADP-ME mutant does not show significant reduction in its Km,NAD values | Homo sapiens |
S346K/K362Q | site-directed mutagenesis | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ATP | 10% inhibition of the wild-type enzyme at 3.0 mM in presence of NAD+, no inhibition in presence of NADP+, inhibition of mutant enzymes by ATP inpresence of NAD+ or NADP+ | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | human m-NAD(P)-ME is a non-cooperative enzyme for substrate L-malate binding, steady-state kinetics of wild-type and mutant enzymes, overview | Homo sapiens | |
0.002 | - |
NADP+ | mutant E314A, pH 7.4, 30°C | Homo sapiens | |
0.003 | - |
NADP+ | mutant E314A/S346K, pH 7.4, 30°C | Homo sapiens | |
0.005 | - |
NADP+ | wild-type enzyme, pH 7.4, 30°C | Homo sapiens | |
0.013 | - |
NADP+ | mutant S346K, pH 7.4, 30°C | Homo sapiens | |
0.03 | - |
NADP+ | mutant K347Y, pH 7.4, 30°C | Homo sapiens | |
0.14 | - |
NADP+ | mutant S346K/K347Y, pH 7.4, 30°C | Homo sapiens | |
0.36 | - |
NADP+ | mutant K362H, pH 7.4, 30°C | Homo sapiens | |
0.4 | - |
(S)-malate | mutant E314A, with NADP+, pH 7.4, 30°C | Homo sapiens | |
0.76 | - |
NADP+ | mutant K362Q, pH 7.4, 30°C | Homo sapiens | |
0.9 | - |
(S)-malate | mutant S346K, with NADP+, pH 7.4, 30°C | Homo sapiens | |
0.9 | - |
NAD+ | mutants E314A/S346K/K347Y/K362H and E314A/S346I/K347D/K362H, pH 7.4, 30°C | Homo sapiens | |
1 | - |
(S)-malate | mutant E314A/S346K/K347Y/K362H, with NAD+, pH 7.4, 30°C | Homo sapiens | |
1 | - |
(S)-malate | mutant S346K/K347Y, with NADP+, pH 7.4, 30°C | Homo sapiens | |
1 | - |
(S)-malate | with NADP+, wild-type enzyme, pH 7.4, 30°C | Homo sapiens | |
1.2 | - |
(S)-malate | mutant E314A/S346K, with NADP+, pH 7.4, 30°C | Homo sapiens | |
1.5 | - |
NAD+ | mutant E314A/S346K/K347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
1.6 | - |
NAD+ | mutant E314A, pH 7.4, 30°C | Homo sapiens | |
1.8 | - |
(S)-malate | mutants K347Y and K362Q, with NADP+, pH 7.4, 30°C | Homo sapiens | |
2 | - |
(S)-malate | mutant E314A/S346K/K347Y/K362Q, with NAD+, pH 7.4, 30°C | Homo sapiens | |
3 | - |
(S)-malate | mutant E314A/S346I/K347D/K362H, with NAD+, pH 7.4, 30°C | Homo sapiens | |
3 | - |
NADP+ | mutant E314A/S346K/K347Y/K362H, pH 7.4, 30°C | Homo sapiens | |
3.7 | - |
(S)-malate | mutant S346K/K362Q, with NADP+, pH 7.4, 30°C | Homo sapiens | |
4 | - |
(S)-malate | mutants E314A, S346K, E314A/S346K, S346K/K362Q, and S346K/K347Y/K362Q, with NAD+, pH 7.4, 30°C | Homo sapiens | |
4.9 | - |
(S)-malate | mutant K362H, with NADP+, pH 7.4, 30°C | Homo sapiens | |
5 | - |
NAD+ | mutant E314A/S346K, pH 7.4, 30°C | Homo sapiens | |
5 | - |
NADP+ | mutant E314A/S346K/K347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
5 | - |
(S)-malate | mutants K347Y, K362Q, and S346K/K347Y, with NAD+, pH 7.4, 30°C | Homo sapiens | |
5 | - |
(S)-malate | with NAD+, wild-type enzyme, pH 7.4, 30°C | Homo sapiens | |
5.5 | - |
(S)-malate | mutant S347Y/K362Q, with NADP+, pH 7.4, 30°C | Homo sapiens | |
5.6 | - |
(S)-malate | mutant E314A/S346K/K347Y/K362Q, with NADP+, pH 7.4, 30°C | Homo sapiens | |
6 | - |
NADP+ | mutant S346K/K362Q, pH 7.4, 30°C | Homo sapiens | |
6 | - |
(S)-malate | mutants S346K/K347Y/K362H, with NAD+, pH 7.4, 30°C | Homo sapiens | |
6.5 | - |
NAD+ | mutant S346I/K347D/K362H, pH 7.4, 30°C | Homo sapiens | |
7 | - |
(S)-malate | mutant S346I/K347D/K362H, with NAD+, pH 7.4, 30°C | Homo sapiens | |
7.1 | - |
NAD+ | mutant S346K/K347Y/K362H, pH 7.4, 30°C | Homo sapiens | |
7.3 | - |
(S)-malate | mutant E314A/S346K/K347Y/K362H, with NADP+, pH 7.4, 30°C | Homo sapiens | |
8 | - |
(S)-malate | mutants K362H and S347Y/K362Q, with NAD+, pH 7.4, 30°C | Homo sapiens | |
10 | - |
NAD+ | mutant S346K/K347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
10 | - |
(S)-malate | mutant S346K/K347Y/K362Q, with NADP+, pH 7.4, 30°C | Homo sapiens | |
11 | - |
NAD+ | mutant K347Y, pH 7.4, 30°C | Homo sapiens | |
11 | - |
(S)-malate | mutant S346K/K347Y/K362H, with NADP+, pH 7.4, 30°C | Homo sapiens | |
12 | - |
NADP+ | mutant S347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
13 | - |
NAD+ | mutant K362Q, pH 7.4, 30°C | Homo sapiens | |
14 | - |
NAD+ | mutant K362H, pH 7.4, 30°C | Homo sapiens | |
14 | - |
NAD+ | mutant S346K/K362Q, pH 7.4, 30°C | Homo sapiens | |
17 | - |
NAD+ | mutant S346K, pH 7.4, 30°C | Homo sapiens | |
17 | - |
NADP+ | mutant S346K/K347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
18 | - |
NAD+ | mutant S346K/K347Y, pH 7.4, 30°C | Homo sapiens | |
18.35 | - |
NAD+ | wild-type enzyme, pH 7.4, 30°C | Homo sapiens | |
20 | - |
NAD+ | mutant S347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
24 | - |
(S)-malate | mutant E314A/S346I/K347D/K362H, with NADP+, pH 7.4, 30°C | Homo sapiens | |
29 | - |
NADP+ | mutants S346K/K347Y/K362H and E314A/S346I/K347D/K362H, pH 7.4, 30°C | Homo sapiens | |
36 | - |
(S)-malate | mutant S346I/K347D/K362H, with NADP+, pH 7.4, 30°C | Homo sapiens | |
116 | - |
NADP+ | mutant S346I/K347D/K362H, pH 7.4, 30°C | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Homo sapiens | 5829 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NADP+ | Homo sapiens | - |
pyruvate + CO2 + NADPH | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD+ | - |
Homo sapiens | pyruvate + CO2 + NADH | - |
r | |
(S)-malate + NADP+ | - |
Homo sapiens | pyruvate + CO2 + NADPH | - |
r |
Synonyms | Comment | Organism |
---|---|---|
c-NADP-ME | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.3 | - |
NADP+ | mutant S346I/K347D/K362H, pH 7.4, 30°C | Homo sapiens | |
8 | - |
NADP+ | mutant S346K/K347Y/K362H, pH 7.4, 30°C | Homo sapiens | |
15 | - |
NADP+ | mutant E314A/S346I/K347D/K362H, pH 7.4, 30°C | Homo sapiens | |
15 | - |
NADP+ | mutant S346K/K347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
22 | - |
NAD+ | mutant S346K, pH 7.4, 30°C | Homo sapiens | |
26 | - |
NADP+ | mutant E314A/S346K/K347Y/K362H, pH 7.4, 30°C | Homo sapiens | |
40 | - |
NAD+ | mutant K347Y, pH 7.4, 30°C | Homo sapiens | |
51 | - |
NADP+ | mutant E314A/S346K/K347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
51 | - |
NAD+ | wild-type enzyme, pH 7.4, 30°C | Homo sapiens | |
53 | - |
NAD+ | mutant S347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
60 | - |
NAD+ | mutant K362Q, pH 7.4, 30°C | Homo sapiens | |
65 | - |
NAD+ | mutant S346K/K347Y, pH 7.4, 30°C | Homo sapiens | |
78 | - |
NADP+ | mutant S347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
102 | - |
NADP+ | mutant S346K/K347Y, pH 7.4, 30°C | Homo sapiens | |
108 | - |
NAD+ | mutant K362H, pH 7.4, 30°C | Homo sapiens | |
110 | - |
NADP+ | mutant E314A, pH 7.4, 30°C | Homo sapiens | |
110 | - |
NAD+ | mutant S346K/K362Q, pH 7.4, 30°C | Homo sapiens | |
112 | - |
NADP+ | mutant S346K/K362Q, pH 7.4, 30°C | Homo sapiens | |
113 | - |
NAD+ | mutant E314A, pH 7.4, 30°C | Homo sapiens | |
121 | - |
NADP+ | mutant K347Y, pH 7.4, 30°C | Homo sapiens | |
124 | - |
NAD+ | mutant S346K/K347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
126 | - |
NADP+ | wild-type enzyme, pH 7.4, 30°C | Homo sapiens | |
129 | - |
NADP+ | mutant S346K, pH 7.4, 30°C | Homo sapiens | |
131 | - |
NAD+ | mutant S346I/K347D/K362H, pH 7.4, 30°C | Homo sapiens | |
132 | - |
NADP+ | mutant K362Q, pH 7.4, 30°C | Homo sapiens | |
137 | - |
NADP+ | mutant E314A/S346K, pH 7.4, 30°C | Homo sapiens | |
145 | - |
NAD+ | mutant E314A/S346K, pH 7.4, 30°C | Homo sapiens | |
149 | - |
NADP+ | mutant K362H, pH 7.4, 30°C | Homo sapiens | |
166 | - |
NAD+ | mutant E314A/S346I/K347D/K362H, pH 7.4, 30°C | Homo sapiens | |
208 | - |
NAD+ | mutant E314A/S346K/K347Y/K362Q, pH 7.4, 30°C | Homo sapiens | |
219 | - |
NAD+ | mutant S346K/K347Y/K362H, pH 7.4, 30°C | Homo sapiens | |
258 | - |
NAD+ | mutant E314A/S346K/K347Y/K362H, pH 7.4, 30°C | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | nucleotide-binding site of c-NADP-ME, sequence comparisons, overview. A series of E314A-containing c-NADP-ME quadruple mutants are changed to NAD+-utilizing enzymes by abrogating NADP+ binding and increasing NAD+ binding | Homo sapiens | |
NAD+ | poor activity with NAD+ compared to NADP+ | Homo sapiens | |
NADP+ | Lys362 is the key residue contributing to binding the 2'-phosphate of NADP+ | Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.73 | - |
ATP | mutants E314A and E314A/S346K, in presence of NAD+, pH 7.4, 30°C | Homo sapiens | |
3.3 | - |
ATP | mutant E314A/S346K/K347Y/K362Q, in presence of NAD+, pH 7.4, 30°C | Homo sapiens | |
3.9 | - |
ATP | mutant E314A/S346K/K347Y/K362Q, in presence of NADP+, pH 7.4, 30°C | Homo sapiens | |
17.2 | - |
ATP | mutant E314A/S346K, in presence of NAPD+, pH 7.4, 30°C | Homo sapiens | |
18.1 | - |
ATP | mutant E314A, in presence of NAPD+, pH 7.4, 30°C | Homo sapiens | |
20.6 | - |
ATP | wild-type enzyme, in presence of NADP+, pH 7.4, 30°C | Homo sapiens | |
23.2 | - |
ATP | wild-type enzyme, in presence of NAD+, pH 7.4, 30°C | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.8 | - |
NAD+ | wild-type enzyme, pH 7.4, 30°C | Homo sapiens | |
36000 | - |
NADP+ | wild-type enzyme, pH 7.4, 30°C | Homo sapiens |