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Literature summary for 2.6.1.44 extracted from

  • Cellini, B.; Bertoldi, M.; Montioli, R.; Paiardini, A.; Borri Voltattorni, C.
    Human wild-type alanine:glyoxylate aminotransferase and its naturally occurring G82E variant: functional properties and physiological implications (2007), Biochem. J., 408, 39-50.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Homo sapiens
wild-type and G82E mutant His-tagged enzyme expressed in Escherichia coli Homo sapiens

Protein Variants

Protein Variants Comment Organism
G82E natural enzyme variant. Significant reduction in affinty for pyridoxal 5'-phosphate and pyridoxamine 5'-phosphate. Mutant displays an altered conformational state of the bound pyridoxal 5'-phosphate and a decrease in overall catlytic activity to 0.1% of wild-type Homo sapiens
G82E naturally occuring variant. Like the wild-type, the G82E variant is able to bind 2 mol pyridoxal 5'-phosphate/dimer, it exhibits a significant reduced affinity for pyridoxal 5'-phosphate and even more for pyridoxamine 5'-phosphate compared with wild-type, and an altered conformational state of the bound pyridoxal 5'-phosphate. Dramatic decrease of the overall catalytic activity (about 0.1% of that of normal alanine:glyoxylate aminotransferase), appears to be related to the inability to undergo an efficient transaldimination of the pyridoxal 5'-phosphate form of the enzyme with amino acids as well as an efficient conversion of AGT-pyridoxamine 5'-phosphate into AGT-pyridoxal 5'-phosphate Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
D-alanine competitive inhibitor with L-alanine as varied substrate and glyoxylate as fixed substrate. Uncompetitive inhibitor with glyoxylate as varied substrate and L-alanine as fixed substrate; competitive with L-alanine, uncompetitive with glyoxylate Homo sapiens
pyruvate mixed type inhibition with L-alanine, competitive with glyoxylate; mixed type inhibitor with L-alanine as varied substrate and glyoxylate as fixed substrate. Competitive inhibitor with glyoxylate as varied substrate and L-alanine as fixed substrate Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.15
-
glyoxylate mutant G82E, 25°C Homo sapiens
0.15
-
glyoxylate pH 7.4, 25°C, mutant enzyme G82E Homo sapiens
0.21
-
pyruvate pH 7.4, 25°C, wild-type enzyme Homo sapiens
0.21
-
pyruvate wild-type, 25°C Homo sapiens
0.23
-
glyoxylate pH 7.4, 25°C, wild-type enzyme Homo sapiens
0.23
-
glyoxylate wild-type, 25°C Homo sapiens
15
-
L-alanine mutant G82E, 25°C Homo sapiens
15
-
L-alanine pH 7.4, 25°C, mutant enzyme G82E Homo sapiens
22
-
glycine pH 7.4, 25°C, wild-type enzyme Homo sapiens
22
-
glycine wild-type, 25°C Homo sapiens
31
-
L-alanine pH 7.4, 25°C, wild-type enzyme Homo sapiens
31
-
L-alanine wild-type, 25°C Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-alanine + glyoxylate Homo sapiens the enzyme is highly specific for catalysing glyoxylate to glycine processing, thereby playing a key role in glyoxylate detoxification pyruvate + glycine
-
?
additional information Homo sapiens enzyme is highly specific for catalyzing glyoxylate to glycine processing, playing a key role in glyoxylate detoxification ?
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-
Homo sapiens A2V838
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and G82E mutant His-tagged enzyme Homo sapiens

Reaction

Reaction Comment Organism Reaction ID
L-alanine + glyoxylate = pyruvate + glycine pyridoxamine 5'-phosphate remains bound to the enzyme during the catalytic cycle. The enzyme-pyridoxamine 5'-phosphate complex displays a reactivity towards oxo acids higher than that of the apo-enzyme in presence of pyridoxamine Homo sapiens

Storage Stability

Storage Stability Organism
-20°C, wild-type and mutant enzyme G82E are stable for at least 1 month Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
glycine + pyruvate
-
Homo sapiens L-alanine + glyoxylate
-
?
L-alanine + glyoxylate the enzyme is highly specific for catalysing glyoxylate to glycine processing, thereby playing a key role in glyoxylate detoxification Homo sapiens pyruvate + glycine
-
?
L-alanine + glyoxylate the enzyme is highly specific for catalysing glyoxylate to glycine processing Homo sapiens pyruvate + glycine
-
?
L-alanine + glyoxylate transamination half-reaction kinetic parameters Homo sapiens glycine + pyruvate
-
r
L-arginine + pyruvate
-
Homo sapiens 5-guanidino-2-oxopentanoate + L-alanine
-
?
L-asparagine + glyoxylate
-
Homo sapiens 4-amino-2,4-dioxobutanoate + glycine
-
?
L-glutamate + glyoxylate
-
Homo sapiens 2-oxoglutarate + glycine
-
?
L-phenylalanine + glyoxylate
-
Homo sapiens phenylpyruvate + glycine
-
?
L-serine + glyoxylate
-
Homo sapiens 3-hydroxy-2-oxopropanoate + glycine
-
?
additional information enzyme is highly specific for catalyzing glyoxylate to glycine processing, playing a key role in glyoxylate detoxification Homo sapiens ?
-
?

Synonyms

Synonyms Comment Organism
AGT
-
Homo sapiens
alanine:glyoxylate aminotransferase
-
Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.068
-
glyoxylate mutant G82E, 25°C Homo sapiens
0.068
-
glyoxylate pH 7.4, 25°C, mutant enzyme G82E Homo sapiens
0.07
-
L-alanine mutant G82E, 25°C Homo sapiens
0.07
-
L-alanine pH 7.4, 25°C, mutant enzyme G82E Homo sapiens
0.33
-
glycine pH 7.4, 25°C, wild-type enzyme Homo sapiens
0.33
-
glycine wild-type, 25°C Homo sapiens
0.36
-
pyruvate pH 7.4, 25°C, wild-type enzyme Homo sapiens
0.36
-
pyruvate wild-type, 25°C Homo sapiens
45
-
glyoxylate pH 7.4, 25°C, wild-type enzyme Homo sapiens
45
-
L-alanine pH 7.4, 25°C, wild-type enzyme Homo sapiens
45
-
glyoxylate wild-type, 25°C Homo sapiens
45
-
L-alanine wild-type, 25°C Homo sapiens

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate like the wild-type, the G82E variant is able to bind 2 mol pyridoxal 5'-phosphate/dimer, it exhibits a significant reduced affinity for pyridoxal 5'-phosphate and even more for pyridoxamine 5'-phosphate compared with wild-type, and an altered conformational state of the bound pyridoxal 5'-phosphate Homo sapiens

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.3
-
pyruvate glyoxylate as varied substrate and L-alanine as fixed substrate Homo sapiens
14.3
-
D-alanine with L-alanine as varied substrate and glyoxylate as fixed substrate, Kis Homo sapiens
15.8
-
pyruvate with L-alanine as varied substrate and glyoxylate as fixed substrate, Kis Homo sapiens
22.8
-
pyruvate with L-alanine as varied substrate and glyoxylate as fixed substrate, Kii Homo sapiens
28.7
-
D-alanine with glyoxylate as varied substrate and L-alanine as fixed substrate, Kii Homo sapiens