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Literature summary extracted from

  • Clark, S.M.; Di Leo, R.; Dhanoa, P.K.; Van Cauwenberghe, O.R.; Mullen, R.T.; Shelp, B.J.
    Biochemical characterization, mitochondrial localization, expression, and potential functions for an Arabidopsis gamma-aminobutyrate transaminase that utilizes both pyruvate and glyoxylate (2009), J. Exp. Bot., 60, 1743-1757.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.6.1.19 co-expression of the truncated GABA-T cDNA with the GroES/ELmolecular chaperone complex in Escherichia coli BL-21(DE3) Rosetta (pLysS) cells, and GABA-T is expressed in Nicotiana tabacum BY-2 cells Arabidopsis thaliana
2.6.1.96 gene pop2, expression of N-termminally His6-tagged GABA-T POP2 in Escherichia coli strain BL-21(DE3) Rosetta (pLysS) Arabidopsis thaliana
2.6.1.96 transient expression of GABA-T in Nicotiana tabacum BY-2 cells. C-terminal myc-tagged GABA-T co-localizes with coexpressed mitochondrial betaATPase-CAT in BY-2 cells Arabidopsis thaliana

Protein Variants

EC Number Protein Variants Comment Organism
2.6.1.96 additional information a truncated AtGABA-T cDNA lacking the putative N-terminal targeting pre-sequence is co-expressed with the molecular chaperones GroES/EL in Escherichia coli, substrate specificity and kinetic studies reveal that the recombinant enzyme utilizes only GABA as the amino donor in the production of succinate semialdehyde Arabidopsis thaliana

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.6.1.19 beta-Alanine competitive inhibitor of pyruvate-dependent GABA-T activity Arabidopsis thaliana
2.6.1.19 glycine competitive inhibitor of pyruvate-dependent GABA-T activity Arabidopsis thaliana
2.6.1.19 ornithine competitive inhibitor of pyruvate-dependent GABA-T activity Arabidopsis thaliana
2.6.1.19 vigabatrin competitive inhibitor of pyruvate-dependent GABA-T activity Arabidopsis thaliana
2.6.1.96 beta-Alanine
-
 Arabidopsis thaliana
2.6.1.96 glycine
-
 Arabidopsis thaliana
2.6.1.96 ornithine
-
 Arabidopsis thaliana
2.6.1.96 vigabatrin
-
 Arabidopsis thaliana

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.6.1.19 0.014
-
 4-Oxobutanoate with L-alanine as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 0.11
-
 glyoxylate in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 0.14
-
 pyruvate in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 0.18
-
 4-aminobutanoate with glyoxylate as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 0.34
-
 4-aminobutanoate with pyruvate as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 2.4
-
 L-alanine with 4-oxobutanoate as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.96 additional information
-
 additional information Michaelis-Menten kinetic analysis, overview Arabidopsis thaliana
2.6.1.96 0.014
-
 succinate semialdehyde reverse reaction with L-alanine, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 0.11
-
 glyoxylate forward reaction with 4-aminobutanoate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 0.14
-
 pyruvate forward reaction with 4-aminobutanoate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 0.14
-
 glyoxylate reverse reaction with L-alanine, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 0.18
-
 4-aminobutanoate forward reaction with glyoxylate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 0.34
-
 4-aminobutanoate forward reaction with pyruvate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 2.2
-
 L-alanine reverse reaction with glyoxylate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 2.4
-
 L-alanine reverse reaction with succinate semialdehyde, pH 9.0, 30°C Arabidopsis thaliana

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.6.1.19 mitochondrion
-
 Arabidopsis thaliana 5739
-
2.6.1.96 mitochondrion
-
 Arabidopsis thaliana 5739
-
2.6.1.96 additional information subcellular localization analysis, overview Arabidopsis thaliana
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.6.1.96 53400
-
 x * 57500, recombinant full-length GABA-T, SDS-PAGE, x * 53400, recombinant truncated GABA-T, SDS-PAGE Arabidopsis thaliana
2.6.1.96 57500
-
 x * 57500, recombinant full-length GABA-T, SDS-PAGE, x * 53400, recombinant truncated GABA-T, SDS-PAGE Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.6.1.96 4-aminobutanoate + glyoxylate Arabidopsis thaliana
-
 succinate semialdehyde + glycine
-
 ir
2.6.1.96 4-aminobutanoate + glyoxylate Arabidopsis thaliana Wassilewskija
-
 succinate semialdehyde + glycine
-
 ir
2.6.1.96 4-aminobutanoate + glyoxylate Arabidopsis thaliana Col-0
-
 succinate semialdehyde + glycine
-
 ir
2.6.1.96 4-aminobutanoate + pyruvate Arabidopsis thaliana
-
 succinate semialdehyde + L-alanine
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.6.1.19 Arabidopsis thaliana Q94CE5 subunit
-
2.6.1.96 Arabidopsis thaliana Q94CE5 gene GABAT1-1 (Salk_007661) and knockout line of ecotype Columbia
-
2.6.1.96 Arabidopsis thaliana Q94CE5 POP2; gene pop2, POP2-3 (CS6387) and knockout line of ecotypes Wassilewskija
-
2.6.1.96 Arabidopsis thaliana Col-0 Q94CE5 gene GABAT1-1 (Salk_007661) and knockout line of ecotype Columbia
-
2.6.1.96 Arabidopsis thaliana Wassilewskija Q94CE5 POP2; gene pop2, POP2-3 (CS6387) and knockout line of ecotypes Wassilewskija
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.6.1.19 ProBond nickel resin column chromatography Arabidopsis thaliana
2.6.1.96 recombinant His6-tagged GABA-T POP2 from Escherichia coli strain BL-21(DE3) Rosetta (pLysS) by nickel affinity chromatography Arabidopsis thaliana

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.6.1.19 root
-
 Arabidopsis thaliana
-
2.6.1.96 seedling
-
 Arabidopsis thaliana
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.6.1.96 additional information
-
 native GABA-T activity in cell-free extracts of Arabidopsis thaliana ecotype Columbia and corresponding knockout mutant, overview Arabidopsis thaliana
2.6.1.96 additional information
-
 native GABA-T activity in cell-free extracts of Arabidopsis thaliana ecotype Wassilewskija and corresponding knockout mutant, overview Arabidopsis thaliana
2.6.1.96 0.0004
-
 crude enzyme extract, substrate pyruvate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 3.53
-
 purified recombinant full-length enzyme, substrate pyruvate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 7.54
-
 purified recombinant truncated enzyme, substrate pyruvate, pH 9.0, 30°C Arabidopsis thaliana

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.6.1.19 4-aminobutanoate + glyoxylate
-
 Arabidopsis thaliana 4-oxobutanoate + glycine
-
 ?
2.6.1.19 4-aminobutanoate + glyoxylate
-
 Arabidopsis thaliana Wassilewskija 4-oxobutanoate + glycine
-
 ?
2.6.1.19 4-aminobutanoate + glyoxylate
-
 Arabidopsis thaliana Col-0 4-oxobutanoate + glycine
-
 ?
2.6.1.19 4-aminobutanoate + pyruvate
-
 Arabidopsis thaliana 4-oxobutanoate + L-alanine
-
 ?
2.6.1.19 4-aminobutanoate + pyruvate
-
 Arabidopsis thaliana Wassilewskija 4-oxobutanoate + L-alanine
-
 ?
2.6.1.19 4-aminobutanoate + pyruvate
-
 Arabidopsis thaliana Col-0 4-oxobutanoate + L-alanine
-
 ?
2.6.1.19 additional information cannot utilize 2-oxoglutarate as amino acceptor Arabidopsis thaliana ?
-
 ?
2.6.1.19 additional information cannot utilize 2-oxoglutarate as amino acceptor Arabidopsis thaliana Wassilewskija ?
-
 ?
2.6.1.19 additional information cannot utilize 2-oxoglutarate as amino acceptor Arabidopsis thaliana Col-0 ?
-
 ?
2.6.1.96 4-aminobutanoate + glyoxylate
-
 Arabidopsis thaliana succinate semialdehyde + glycine
-
 ir
2.6.1.96 4-aminobutanoate + glyoxylate the glyoxylate-dependent reaction is not reversible, although glyoxylate can also be used in conjunction with alanine as a substrate in the reverse reaction Arabidopsis thaliana succinate semialdehyde + glycine
-
 ir
2.6.1.96 4-aminobutanoate + glyoxylate
-
 Arabidopsis thaliana Wassilewskija succinate semialdehyde + glycine
-
 ir
2.6.1.96 4-aminobutanoate + glyoxylate
-
 Arabidopsis thaliana Col-0 succinate semialdehyde + glycine
-
 ir
2.6.1.96 4-aminobutanoate + pyruvate
-
 Arabidopsis thaliana succinate semialdehyde + L-alanine
-
 r
2.6.1.96 4-aminobutanoate + pyruvate the pyruvate-dependent GABA-T reaction is reversible Arabidopsis thaliana succinate semialdehyde + L-alanine
-
 r
2.6.1.96 glyoxylate + L-alanine
-
 Arabidopsis thaliana ?
-
 ?
2.6.1.96 additional information GABA-T catalyses the breakdown of GABA to succinic semialdehyde. The enzyme shows both pyruvate- and glyoxylate-dependent activities, but no activity with 2-oxoglutarate. The substrate specificity at pH 9 reveals that the enzyme utilizes glyoxylate, as well as pyruvate, as amino acceptors in the forward reaction, and GABA, but not beta-alanine, ornithine, acetylornithine, serine, glycine, asparagine, glutamine, glutamate, valine, leucine, isoleucine, methionine, phenylalanine, histidine, lysine, arginine, aspartate, threonine, tyrosine, tryptophan, proline, or cysteine as amino donors Arabidopsis thaliana ?
-
 ?
2.6.1.96 additional information GABA-T catalyses the breakdown of GABA to succinic semialdehyde. The enzyme shows both pyruvate- and glyoxylate-dependent activities, but no activity with 2-oxoglutarate. The substrate specificity at pH 9 reveals that the enzyme utilizes glyoxylate, as well as pyruvate, as amino acceptors in the forward reaction, and GABA, but not beta-alanine, ornithine, acetylornithine, serine, glycine, asparagine, glutamine, glutamate, valine, leucine, isoleucine, methionine, phenylalanine, histidine, lysine, arginine, aspartate, threonine, tyrosine, tryptophan, proline, or cysteine as amino donors Arabidopsis thaliana Wassilewskija ?
-
 ?
2.6.1.96 additional information GABA-T catalyses the breakdown of GABA to succinic semialdehyde. The enzyme shows both pyruvate- and glyoxylate-dependent activities, but no activity with 2-oxoglutarate. The substrate specificity at pH 9 reveals that the enzyme utilizes glyoxylate, as well as pyruvate, as amino acceptors in the forward reaction, and GABA, but not beta-alanine, ornithine, acetylornithine, serine, glycine, asparagine, glutamine, glutamate, valine, leucine, isoleucine, methionine, phenylalanine, histidine, lysine, arginine, aspartate, threonine, tyrosine, tryptophan, proline, or cysteine as amino donors Arabidopsis thaliana Col-0 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.6.1.96 ? x * 57500, recombinant full-length GABA-T, SDS-PAGE, x * 53400, recombinant truncated GABA-T, SDS-PAGE Arabidopsis thaliana

Synonyms

EC Number Synonyms Comment Organism
2.6.1.19 GABA-T
-
 Arabidopsis thaliana
2.6.1.19 gamma-aminobutyrate transaminase
-
 Arabidopsis thaliana
2.6.1.96 GABA-T
-
 Arabidopsis thaliana
2.6.1.96 gamma-aminobutyrate transaminase
-
 Arabidopsis thaliana
2.6.1.96 Pop2
-
 Arabidopsis thaliana

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.6.1.96 30
-
 assay at Arabidopsis thaliana

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.6.1.19 6.9
-
 4-aminobutanoate with glyoxylate as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 9.7
-
 glyoxylate in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 10.6
-
 pyruvate in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 10.6
-
 4-aminobutanoate with pyruvate as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 10.8
-
 4-Oxobutanoate with L-alanine as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 15.4
-
 L-alanine with 4-oxobutanoate as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.96 6.9
-
 4-aminobutanoate forward reaction with glyoxylate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 9.7
-
 glyoxylate forward reaction with 4-aminobutanoate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 10.6
-
 pyruvate forward reaction with 4-aminobutanoate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 10.6
-
 4-aminobutanoate forward reaction with pyruvate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 10.8
-
 succinate semialdehyde reverse reaction with L-alanine, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 12.1
-
 glyoxylate reverse reaction with L-alanine, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 14.1
-
 L-alanine reverse reaction with glyoxylate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 15.4
-
 L-alanine reverse reaction with succinate semialdehyde, pH 9.0, 30°C Arabidopsis thaliana

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.6.1.19 9
-
 for pyruvate-dependent GABA-T activity Arabidopsis thaliana
2.6.1.96 9
-
 pyruvate-dependent forward reaction Arabidopsis thaliana

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.6.1.19 8.5 10
-
 Arabidopsis thaliana
2.6.1.96 8 10.5 pyruvate-dependent forward reaction Arabidopsis thaliana

Cofactor

EC Number Cofactor Comment Organism Structure
2.6.1.19 pyridoxal 5'-phosphate
-
 Arabidopsis thaliana
2.6.1.96 pyridoxal 5'-phosphate
-
 Arabidopsis thaliana

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.6.1.19 0.46
-
 ornithine
-
 Arabidopsis thaliana
2.6.1.19 0.55
-
 beta-Alanine
-
 Arabidopsis thaliana
2.6.1.19 0.62
-
 vigabatrin
-
 Arabidopsis thaliana
2.6.1.19 3.7
-
 glycine
-
 Arabidopsis thaliana
2.6.1.96 0.46
-
 ornithine forward reaction with 4-aminobutanoate and pyruvate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 0.55
-
 beta-Alanine forward reaction with 4-aminobutanoate and pyruvate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 0.62
-
 vigabatrin forward reaction with 4-aminobutanoate and pyruvate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 3.7
-
 glycine forward reaction with 4-aminobutanoate and pyruvate, pH 9.0, 30°C Arabidopsis thaliana

Expression

EC Number Organism Comment Expression
2.6.1.19 Arabidopsis thaliana expression is highest in roots and increases as a function of leaf development up

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.6.1.19 6.4
-
 L-alanine with 4-oxobutanoate as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 31
-
 4-aminobutanoate with pyruvate as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 39
-
 4-aminobutanoate with glyoxylate as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 76
-
 pyruvate in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 88
-
 glyoxylate in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.19 770
-
 4-Oxobutanoate with L-alanine as cosubstrate, in 50 mM TABS (pH 9), 1.5 mM dithiothreitol, 0.625 mM EDTA, 0.1 mM pyridoxal 5'-phosphate, 10% (v/v) glycerol, at 30°C Arabidopsis thaliana
2.6.1.96 6.4
-
 L-alanine reverse reaction with succinate semialdehyde, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 6.5
-
 L-alanine reverse reaction with glyoxylate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 31
-
 4-aminobutanoate forward reaction with pyruvate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 39
-
 4-aminobutanoate forward reaction with glyoxylate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 76
-
 pyruvate forward reaction with 4-aminobutanoate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 86
-
 glyoxylate reverse reaction with L-alanine, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 88
-
 glyoxylate forward reaction with 4-aminobutanoate, pH 9.0, 30°C Arabidopsis thaliana
2.6.1.96 770
-
 succinate semialdehyde reverse reaction with L-alanine, pH 9.0, 30°C Arabidopsis thaliana