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Literature summary for 1.4.1.2 extracted from

  • Paradisi, F.; Dean, J.L.; Geoghegan, K.F.; Engel, P.C.
    Spontaneous chemical reversion of an active site mutation: deamidation of an asparagine residue replacing the catalytic aspartic acid of glutamate dehydrogenase (2005), Biochemistry, 44, 3636-3643.
    View publication on PubMed
Search for more literature for 1.4.1.2

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli TG1 cells [Clostridium] symbiosum

Protein Variants

Protein Variants Comment Organism
D165N residual 2% of wild-type activity when purified after expression in Escherichia coli at 37°C, cells induced at 8°C are 1000fold less active than that produced at 37°C, spontaneous deamidation, which depends on the residual catalytic machinery of the mutated GDH active site [Clostridium] symbiosum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0056
-
 NADH mutant D165N [Clostridium] symbiosum
0.0108
-
 NADH wild-type [Clostridium] symbiosum
0.076
-
 NAD+ mutant D165N [Clostridium] symbiosum
0.0862
-
 2-oxoglutarate mutant D165N [Clostridium] symbiosum
0.125
-
 2-oxoglutarate wild-type [Clostridium] symbiosum
0.14
-
 NAD+ wild-type [Clostridium] symbiosum
2.66
-
 L-glutamate mutant D165N [Clostridium] symbiosum
3.97
-
 L-glutamate wild-type [Clostridium] symbiosum
49.4
-
 NH4+ mutant D165N [Clostridium] symbiosum
62.8
-
 NH4+ wild-type [Clostridium] symbiosum

Organism

Organism UniProt Comment Textmining
[Clostridium] symbiosum
-
-
-

Purification (Commentary)

Purification (Comment) Organism
to homogeneity [Clostridium] symbiosum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0003
-
 mutant D165N overexpressed at 8°C [Clostridium] symbiosum
0.034
-
 mutant D165N overexpressed at 37°C [Clostridium] symbiosum
0.047
-
 mutant D165N overexpressed at 23°C [Clostridium] symbiosum

Storage Stability

Storage Stability Organism
-20°C [Clostridium] symbiosum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxoglutarate + NH4+ + NADPH
-
 [Clostridium] symbiosum L-glutamate + NADP+
-
 ?
L-glutamate + NAD+ + H2O
-
 [Clostridium] symbiosum 2-oxoglutarate + NADH + NH3
-
 r

Synonyms

Synonyms Comment Organism
GDH
-
 [Clostridium] symbiosum
glutamate dehydrogenase
-
 [Clostridium] symbiosum

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
8 37 decreasing temperature results in much lower enzyme activity [Clostridium] symbiosum

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 [Clostridium] symbiosum