1.1.1.82: malate dehydrogenase (NADP+)

This is an abbreviated version, for detailed information about malate dehydrogenase (NADP+), go to the full flat file.

Reaction

(S)-malate
+
NADP+
=
oxaloacetate
+
NADPH
+
H+

Synonyms

(S)-malate dehydrogenase, dehydrogenase, malate (nicotinamide adenine dinucleotide phosphate), malate NADP dehydrogenase, malic dehydrogenase (nicotinamide adenine dinucleotide phosphate), MDH, NADP malate dehydrogenase, NADP-dependent malate dehydrogenase, NADP-linked malate dehydrogenase, NADP-malate dehydrogenase, NADP-malic enzyme, NADP-MDH, NADP-MDH1, NADP-MDH2, NADPH-MDH

ECTree

     1 Oxidoreductases
         1.1 Acting on the CH-OH group of donors
             1.1.1 With NAD+ or NADP+ as acceptor
                1.1.1.82 malate dehydrogenase (NADP+)

Engineering

Engineering on EC 1.1.1.82 - malate dehydrogenase (NADP+)

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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C23A
-
minute activity under oxidizing conditions, up to 1300fold activation under reducing conditions
C23A/C28A
-
minute activity under oxidizing conditions, up to 1300fold activation under reducing conditions
C23A/C28A/C206A
-
only marginal activity in the oxidized state, does not exhibit full activity in the crude extract
C28A
-
minute activity under oxidizing conditions, up to 1300fold activation under reducing conditions
DELTAN
-
mutant lacking the N-terminal 45 residues
C207A
Sorghum sp.
-
strictly thioredoxin-dependent, totally insensitive to diethyl dicarbonate
C207A/DETAEV
Sorghum sp.
-
very low spontaneous activity, high Km-value for oxaloacetate, fast activation kinetics in the presence of reduced thioredoxin, ability to be activated by dithiothreitol alone at a slow rate
C207A/E387Q
Sorghum sp.
-
very low spontaneous activity, high Km-value for oxaloacetate, fast activation kinetics in the presence of reduced thioredoxin, ability to be activated by dithiothreitol alone at a slow rate
C24S/C207A
Sorghum sp.
-
oxidation-reduction midpoint potential is -310 mV at pH 7.0, compared to -330 mV for the wild-type enzyme
C24S/C29S
Sorghum sp.
-
oxidation-reduction midpoint potential is -280 mV at pH 7.0, compared to -330 mV for the wild-type enzyme
C29S
Sorghum sp.
-
strictly thioredoxin-dependent, totally insensitive to diethyl dicarbonate
C29S/C207A
Sorghum sp.
-
no spontaneous activity, activated much faster than the wild-type protein, strictly dependent on reduced thioredoxin for activation
C29S/C207A/DELTAEV
Sorghum sp.
-
high spontaneous activity, activated by reduced thioredoxin almost instantaneously and also by dithiothreitol alone, although at a much slower rate. The Km-values for both the oxidized and reduced enzyme show no significant differences in the apparent affinity for NADPH, whereas the Km for oxaloacetate is dramatically increased in the oxidized form
C29S/C207A/E387Q
Sorghum sp.
-
high spontaneous activity, activated by reduced thioredoxin almost instantaneously and also by dithiothreitol alone, although at a much slower rate. The Km-values for both the oxidized and reduced enzyme show no significant differences in the apparent affinity for NADPH, whereas the Km for oxaloacetate is dramatically increased in the oxidized form
C29S/DELTAEV
Sorghum sp.
-
very low spontaneous activity, high Km-value for oxaloacetate, fast activation kinetics in the presence of reduced thioredoxin, ability to be activated by dithiothreitol alone at a slow rate
C39S/E387Q
Sorghum sp.
-
very low spontaneous activity, high Km-value for oxaloacetate, fast activation kinetics in the presence of reduced thioredoxin, ability to be activated by dithiothreitol alone at a slow rate
C64S
Sorghum sp.
-
still requires activation by reduced thioredoxin, activation is almost instantaneous, whereas the native enzyme reaches full activity after 10-20 min of preincubation, the half-saturation concentration for reduced thioredoxin is decreased 2fold
C64S/C69S
Sorghum sp.
-
still requires activation by reduced thioredoxin, activation is almost instantaneous, whereas the native enzyme reaches full activity after 10-20 min of preincubation, the half-saturation concentration for reduced thioredoxin is decreased 2fold
C69S
Sorghum sp.
-
still requires activation by reduced thioredoxin, activation is almost instantaneous, whereas the native enzyme reaches full activity after 10-20 min of preincubation, the half-saturation concentration for reduced thioredoxin is decreased 2fold
D201A
Sorghum sp.
-
only slightly active, 80fold increased Km for oxaloacetate, Km-value for NADPH is slightly decreased
D201N
Sorghum sp.
-
only slightly active, 45fold increased Km for oxaloacetate, Km-value for NADPH is slightly decreased
DELTAEV
Sorghum sp.
-
mutant with the two most C-terminal residues deleted, NADP+ does not inhibit activation, activation time course of thioredoxin-dependent activation of both mutant proteins is similar to that of the wild-type protein
DELTAN
Sorghum sp.
-
truncation mutant corresponding to the deletion of the 5' end of the mdh cDNA open reading frame until the 73rd codon
E387Q
Sorghum sp.
-
NADP+ does not inhibit activation, activation time course of thioredoxin-dependent activation of both mutant proteins is similar to that of the wild-type protein
G84D
Sorghum sp.
-
10fold lower Km for NADH, Km for NADPH remains unchanged
G84D/S851I/R87Q/S88A
Sorghum sp.
-
changed cofactor specificity from NADPH to NADH, the activation of the NAD-specific thiol-regulated enzyme is inhibited by NAD+ but no longer by NADP+
H229N
Sorghum sp.
-
no activity
H229Q
Sorghum sp.
-
no activity
S85I/R87Q/S88A
Sorghum sp.
-
7fold increase in the Km-value for NADPH and 4fold decrease in Km-value for NADH
additional information