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Literature summary for 1.4.1.3 extracted from

  • Fang, J.; Hsu, B.Y.L.; MacMullen, C.M.; Poncz, M.; Smith, T.J.; Stanley, C.A.
    Expression, purification and characterization of human glutamate dehydrogenase (GDH) allosteric regulatory mutations (2002), Biochem. J., 363, 81-87.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
ADP no activation of R463A mutant enzyme Homo sapiens
ATP wild-type: activation at 1 mM, inhibition below and above, H454Y and S448P mutant enzymes: progressive increase in activity until 10 mM, inhibition above Homo sapiens
leucine wild-type, S448P, H454Y and R463A mutant enzymes Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
coexpression of wild-type and mutant enzymes with GroES and GroEl in Escherichia coli Homo sapiens

Protein Variants

Protein Variants Comment Organism
H454Y lower basal activity but comparable maximal activity as wild-type Homo sapiens
R463A stimulatory effect of ADP is eliminated Homo sapiens
S448P unstable in Tris-buffer especially in the absence of allosteric activators, basal and maximal specific activity is lower than that from wild-type Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
ATP wild-type: inhibition at 0.1 mM and between 0.5-1.0 mM and above, activation at 1 mM, H454Y and S448P mutant enzyme: activation between 0.1-1 mM, inhibition above, R463A mutant enzyme: progressive inhibition between 0.01 and 10 mM Homo sapiens
diethylstilbestrol
-
Homo sapiens
GTP little inhibition of H454Y mutant enzyme Homo sapiens
palmitoyl-CoA
-
Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutant enzyme, Q-Sepharose, omega-aminopentyl column, GTP-agarose Homo sapiens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [Āµmol/min/mg] Specific Activity Maximum [Āµmol/min/mg] Comment Organism
19
-
recombinant S448P mutant enzyme, basal activity Homo sapiens
55
-
recombinant S448P mutant enzyme, maximal activity in the presence of 6 mM leucine Homo sapiens
69
-
recombinant S448P mutant enzyme, maximal activity in the presence of 0.2 mM ADP Homo sapiens
73
-
recombinant R463A mutant enzyme, basal activity Homo sapiens
74
-
recombinant H454Y mutant enzyme, basal activity Homo sapiens
84
-
recombinant wild-type enzyme, basal activity Homo sapiens
100
-
recombinant R463A mutant enzyme, maximal activity in the presence of 6 mM leucine Homo sapiens
130
-
recombinant wild-type enzyme, maximal activity in the presence of 0.2 mM ADP or 6 mM leucine Homo sapiens
130
-
recombinant S448P mutant enzyme, maximal activity in the presence of 6 mM leucine Homo sapiens
140
-
recombinant H454Y mutant enzyme, maximal activity in the presence of 0.2 mM ADP Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-glutamate + H2O + NAD(P)+
-
Homo sapiens 2-oxoglutarate + NH3 + NAD(P)H
-
r

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000013
-
palmitoyl-CoA inhibition of recombinant S448P mutant enzyme Homo sapiens
0.000015
-
palmitoyl-CoA inhibition of recombinant H454Y mutant enzyme Homo sapiens
0.00003
-
palmitoyl-CoA inhibition of recombinant wild-type enzyme Homo sapiens
0.000042
-
GTP inhibition of recombinant wild-type enzyme Homo sapiens
0.0002
-
palmitoyl-CoA inhibition of recombinant R463A mutant enzyme Homo sapiens
0.00022
-
GTP inhibition of recombinant R463A mutant enzyme Homo sapiens
0.00036
-
diethylstilbestrol inhibition of recombinant H454Y mutant enzyme Homo sapiens
0.00074
-
diethylstilbestrol inhibition of recombinant S448P mutant enzyme Homo sapiens
0.0017
-
diethylstilbestrol inhibition of recombinant wild-type enzyme Homo sapiens
0.0031
-
GTP inhibition of recombinant S448P mutant enzyme Homo sapiens
0.0051
-
diethylstilbestrol inhibition of recombinant R463A mutant enzyme Homo sapiens
0.21
-
GTP inhibition of recombinant H454Y mutant enzyme Homo sapiens