Literature summary for 2.6.1.78 extracted from

Giustini, C.; Graindorge, M.; Cobessi, D.; Crouzy, S.; Robin, A.; Curien, G.; Matringe, M.
Tyrosine metabolism identification of a key residue in the acquisition of prephenate aminotransferase activity by 1beta aspartate aminotransferase (2019), FEBS J., 286, 2118-2134 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene aat, recombinant expression in Escherichia coli strain BL21 (DE3) Rosetta2	Arabidopsis thaliana
gene aat, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) Rosetta2	Thermus thermophilus
gene aat, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) Rosetta2	Sinorhizobium meliloti

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme, crystallization from 0.1 M Na-citrate, pH 4.0, 11% PEG 4000, 20°C, X-ray diffraction structure determination and analysis at 1.7 A resolution	Arabidopsis thaliana
purified recombinant enzyme, Rme-AAT/PAT crystals are obtained from 0.2 M ammonium formate, 19% PEG 3350, and Rme-AAT grow from 0.1 M Na acetate pH 4.5, 5% PEG 4000, protein concentration is 10 mg/ml, at 20°C, X-ray diffraction structure determination and analysis at 1.79-1.9 A resolution	Sinorhizobium meliloti

Protein Variants

Protein Variants	Comment	Organism
K12G	naturally occuring mutant, the incompetent PATs of Rhizobium meliloti (UniProt ID P58350) has a G instead of K12 and shows reduced PAT activity at prephenate concentrations up to 2.5 mM using a coupled assay method. The mutant shows reduced activity and altered kinetics compared to the wild-type	Sinorhizobium meliloti
K12G	site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type	Thermus thermophilus
additional information	site-directed mutagenesis, modeling, and molecular dynamics simulations reveal that K/R/Q12 residue (numbering according to Thermus thermophilus 1beta AAT) is a signature of the PAT function of 1bta AAT. It is present in the N-terminal flexible loop only in PAT competent 1beta-AAT and has a possible role in stabilizing prephenate by interacting with its 4-hydroxy group	Sinorhizobium meliloti

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	Michaelis-Menten steady-state kinetics analysis	Thermus thermophilus
additional information	-	additional information	Michaelis-Menten steady-state kinetics analysis	Sinorhizobium meliloti
0.0121	-	oxaloacetate	pH 8.0, 30°C, wild-type enzyme	Sinorhizobium meliloti
0.0196	-	oxaloacetate	pH 8.0, 30°C, mutant K12G	Sinorhizobium meliloti
0.0253	-	oxaloacetate	pH 8.0, 30°C, wild-type enzyme	Thermus thermophilus
0.0264	-	oxaloacetate	pH 8.0, 30°C, mutant K12G	Thermus thermophilus
0.114	-	prephenate	pH 8.0, 30°C, wild-type enzyme	Sinorhizobium meliloti
0.15	-	prephenate	pH 8.0, 30°C, wild-type enzyme	Thermus thermophilus
0.632	-	prephenate	pH 8.0, 30°C, mutant K12G	Sinorhizobium meliloti
2.332	-	prephenate	pH 8.0, 30°C, mutant K12G	Thermus thermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
L-arogenate + oxaloacetate	Arabidopsis thaliana	-	L-aspartate + prephenate	-	r
L-arogenate + oxaloacetate	Thermus thermophilus	-	L-aspartate + prephenate	-	r
L-arogenate + oxaloacetate	Sinorhizobium meliloti	-	L-aspartate + prephenate	-	r
L-arogenate + oxaloacetate	Thermus thermophilus DSM 579	-	L-aspartate + prephenate	-	r
L-arogenate + oxaloacetate	Thermus thermophilus ATCC 27634	-	L-aspartate + prephenate	-	r

Organism

Organism	UniProt	Comment	Textmining
Arabidopsis thaliana	Q9SIE1	-	-
Sinorhizobium meliloti	Q02635	-	-
Thermus thermophilus	Q56232	-	-
Thermus thermophilus ATCC 27634	Q56232	-	-
Thermus thermophilus DSM 579	Q56232	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
L-arogenate + oxaloacetate	-	Arabidopsis thaliana	L-aspartate + prephenate	-	r
L-arogenate + oxaloacetate	-	Thermus thermophilus	L-aspartate + prephenate	-	r
L-arogenate + oxaloacetate	-	Sinorhizobium meliloti	L-aspartate + prephenate	-	r
L-arogenate + oxaloacetate	-	Thermus thermophilus DSM 579	L-aspartate + prephenate	-	r
L-arogenate + oxaloacetate	-	Thermus thermophilus ATCC 27634	L-aspartate + prephenate	-	r
L-aspartate + prephenate	-	Thermus thermophilus	L-arogenate + oxaloacetate	-	r
L-aspartate + prephenate	-	Sinorhizobium meliloti	L-arogenate + oxaloacetate	-	r
L-aspartate + prephenate	-	Thermus thermophilus DSM 579	L-arogenate + oxaloacetate	-	r
L-aspartate + prephenate	-	Thermus thermophilus ATCC 27634	L-arogenate + oxaloacetate	-	r

Synonyms

Synonyms	Comment	Organism
1beta AAT	-	Arabidopsis thaliana
1beta AAT	-	Thermus thermophilus
1beta AAT	-	Sinorhizobium meliloti
1beta AAT prephenate aminotransferase	-	Arabidopsis thaliana
1beta AAT prephenate aminotransferase	-	Thermus thermophilus
1beta AAT prephenate aminotransferase	-	Sinorhizobium meliloti
1beta aspartate aminotransferase	-	Arabidopsis thaliana
1beta aspartate aminotransferase	-	Thermus thermophilus
1beta aspartate aminotransferase	-	Sinorhizobium meliloti
aatA	-	Sinorhizobium meliloti
aspartate/prephenate aminotransferase	UniProt	Thermus thermophilus
aspartate/prephenate aminotransferase	UniProt	Sinorhizobium meliloti
aspC	-	Thermus thermophilus
Ath-PAT	-	Arabidopsis thaliana
bifunctional aspartate aminotransferase and glutamate/aspartate-prephenate aminotransferase	UniProt	Arabidopsis thaliana
More	cf. EC 2.6.1.1	Thermus thermophilus
More	cf. EC 2.6.1.79 and EC 2.6.1.1	Arabidopsis thaliana
More	cf. EC 2.6.1.79 and EC 2.6.1.1	Sinorhizobium meliloti
Pat	-	Arabidopsis thaliana
Pat	-	Thermus thermophilus
Pat	-	Sinorhizobium meliloti
Rme-AAT	-	Sinorhizobium meliloti
Tth-PAT	-	Thermus thermophilus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Thermus thermophilus
30	-	assay at	Sinorhizobium meliloti

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
4.8	-	prephenate	pH 8.0, 30°C, mutant K12G	Thermus thermophilus
7.7	-	prephenate	pH 8.0, 30°C, wild-type enzyme	Thermus thermophilus
12.3	-	oxaloacetate	pH 8.0, 30°C, mutant K12G	Thermus thermophilus
32.2	-	oxaloacetate	pH 8.0, 30°C, wild-type enzyme	Thermus thermophilus
38.6	-	prephenate	pH 8.0, 30°C, mutant K12G	Sinorhizobium meliloti
42.3	-	prephenate	pH 8.0, 30°C, wild-type enzyme	Sinorhizobium meliloti
140.2	-	oxaloacetate	pH 8.0, 30°C, mutant K12G	Sinorhizobium meliloti
158.4	-	oxaloacetate	pH 8.0, 30°C, wild-type enzyme	Sinorhizobium meliloti

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Thermus thermophilus
8	-	assay at	Sinorhizobium meliloti

Cofactor

Cofactor	Comment	Organism
pyridoxal 5'-phosphate	-	Arabidopsis thaliana
pyridoxal 5'-phosphate	-	Thermus thermophilus
pyridoxal 5'-phosphate	-	Sinorhizobium meliloti

General Information

General Information	Comment	Organism
malfunction	one PAT incompetent 1beta AAT mutant K12G from Rhizobium meliloti is detected (UniProt ID P58350)	Sinorhizobium meliloti
metabolism	the enzyme is involved in the tyrosine/phenylalanine biosynthesis pathway, overview	Arabidopsis thaliana
metabolism	the enzyme is involved in the tyrosine/phenylalanine biosynthesis pathway, overview	Thermus thermophilus
metabolism	the enzyme is involved in the tyrosine/phenylalanine biosynthesis pathway, overview	Sinorhizobium meliloti
additional information	modeling and molecular dynamic simulations. Identification of a molecular determinant of PAT activity in the flexible N-terminal loop of 1beta AAT. A Lys/Arg/Gln residue in position 12 (numbering according to Thermus thermophilus 1beta AAT), present only in PAT competent enzymes, can interact with the 4-hydroxyl group of the prephenate substrate, probably playing a central role in the acquisition of PAT activity by 1beta AAT. PAT enzyme structures comparisons, overview	Arabidopsis thaliana
additional information	modeling and molecular dynamic simulations. Identification of a molecular determinant of PAT activity in the flexible N-terminal loop of 1beta AAT. A Lys/Arg/Gln residue in position 12 (numbering according to Thermus thermophilus 1beta AAT), present only in PAT competent enzymes, can interact with the 4-hydroxyl group of the prephenate substrate, probably playing a central role in the acquisition of PAT activity by 1beta AAT. PAT enzyme structures comparisons, overview	Thermus thermophilus
additional information	modeling and molecular dynamic simulations. Identification of a molecular determinant of PAT activity in the flexible N-terminal loop of 1beta AAT. A Lys/Arg/Gln residue in position 12 (numbering according to Thermus thermophilus 1beta AAT), present only in PAT competent enzymes, can interact with the 4-hydroxyl group of the prephenate substrate, probably playing a central role in the acquisition of PAT activity by 1beta AAT. PAT enzyme structures comparisons, overview. Lys12 plays a specific role in prephenate binding by PAT competent 1beta AAT	Sinorhizobium meliloti

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
2.06	-	prephenate	pH 8.0, 30°C, mutant K12G	Thermus thermophilus
51.3	-	prephenate	pH 8.0, 30°C, wild-type enzyme	Thermus thermophilus
61.1	-	prephenate	pH 8.0, 30°C, mutant K12G	Sinorhizobium meliloti
371.1	-	prephenate	pH 8.0, 30°C, wild-type enzyme	Sinorhizobium meliloti
465.9	-	oxaloacetate	pH 8.0, 30°C, mutant K12G	Thermus thermophilus
1272.7	-	oxaloacetate	pH 8.0, 30°C, wild-type enzyme	Thermus thermophilus
7153.1	-	oxaloacetate	pH 8.0, 30°C, mutant K12G	Sinorhizobium meliloti
13090.9	-	oxaloacetate	pH 8.0, 30°C, wild-type enzyme	Sinorhizobium meliloti