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Literature summary extracted from

  • Ziegler, K.; Noble, S.M.; Mutumanje, E.; Bishop, B.; Huddler, D.P.; Born, T.L.
    Identification of catalytic cysteine, histidine, and lysine residues in Escherichia coli homoserine transsuccinylase (2007), Biochemistry, 46, 2674-2683.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.46 gene metA, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
2.3.1.46 C142A site-directed mutagenesis, the mutant is inactive Escherichia coli
2.3.1.46 C142S site-directed mutagenesis, the mutant is inactive Escherichia coli
2.3.1.46 C90A site-directed mutagenesis, the mutant shows only slightly reduced activity compared to the wild-type enzyme Escherichia coli
2.3.1.46 C90S site-directed mutagenesis, the mutant shows only slightly reduced activity compared to the wild-type enzyme Escherichia coli
2.3.1.46 H235A site-directed mutagenesis, the mutant is inactive Escherichia coli
2.3.1.46 H235N site-directed mutagenesis, the mutant is inactive Escherichia coli
2.3.1.46 K45A/K46A site-directed mutagenesis, the mutant is nearly inactive Escherichia coli
2.3.1.46 K46A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Escherichia coli
2.3.1.46 K46R site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Escherichia coli
2.3.1.46 K46R/K47R site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Escherichia coli
2.3.1.46 K47A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Escherichia coli
2.3.1.46 K47R site-directed mutagenesis, the mutant shows 90% reduced activity compared to the wild-type enzyme Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.1.46 diethyldicarbonate inactivation, pH dependence, overview Escherichia coli
2.3.1.46 iodoacetamide inactivation, pH dependence, overview Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.46 additional information
-
 additional information steady-state kinetics Escherichia coli
2.3.1.46 0.0032
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K47A Escherichia coli
2.3.1.46 0.027
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46R/K47R Escherichia coli
2.3.1.46 0.044
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46A Escherichia coli
2.3.1.46 0.049
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K47R Escherichia coli
2.3.1.46 0.094
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K47R Escherichia coli
2.3.1.46 0.13
-
 succinyl-CoA pH 7.2, 25°C, recombinant wild-type enzyme Escherichia coli
2.3.1.46 0.13
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46R Escherichia coli
2.3.1.46 0.13
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46R/K47R Escherichia coli
2.3.1.46 0.14
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant C90S Escherichia coli
2.3.1.46 0.14
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46A Escherichia coli
2.3.1.46 0.16
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K47A Escherichia coli
2.3.1.46 0.58
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46R Escherichia coli
2.3.1.46 0.72
-
 L-homoserine pH 7.2, 25°C, recombinant wild-type enzyme Escherichia coli
2.3.1.46 1.07
-
 L-homoserine pH 7.2, 25°C, recombinant mutant C90S Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.1.46 succinyl-CoA + L-homoserine Escherichia coli step in the biosynthetic pathways of methionine and S-adenosylmethionine, overview CoA + O-succinyl-L-homoserine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.46 Escherichia coli
-
 gene metA
-
2.5.1.48 Escherichia coli
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.46 recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by anion exchange chromatography, ammonium sulfate fractionation, hydrophobic interaction chromatography, and dialysis to over 90% purity Escherichia coli

Reaction

EC Number Reaction Comment Organism Reaction ID
2.3.1.46 succinyl-CoA + L-homoserine = CoA + O-succinyl-L-homoserine ping pong catalytic reaction mechanism, the residues Cys142, His235, and Lys47 are essential for catalytic activity, Cys142 acts as nucleophile to which the succinyl moiety is transferred during the reaction followed by transfer to homoserine to form O-succinylhomoserine, Lys47 is involved in the first half-reaction, and His235 is the active site base Escherichia coli
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.46 succinyl-CoA + L-homoserine
-
 Escherichia coli CoA + O-succinyl-L-homoserine
-
 ?
2.3.1.46 succinyl-CoA + L-homoserine step in the biosynthetic pathways of methionine and S-adenosylmethionine, overview Escherichia coli CoA + O-succinyl-L-homoserine
-
 ?
2.5.1.48 O-acetyl-L-homoserine + L-cysteine
-
 Escherichia coli L-cystathionine + acetate
-
 ?
2.5.1.48 O-succinyl-L-homoserine + L-cysteine
-
 Escherichia coli L-cystathionine + succinate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.1.46 homoserine transsuccinylase
-
 Escherichia coli
2.5.1.48 homoserine transsuccinylase
-
 Escherichia coli
2.5.1.48 HTS
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.1.46 25
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.1.46 0.034
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K47A Escherichia coli
2.3.1.46 0.34
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K47A Escherichia coli
2.3.1.46 1.8
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K47R Escherichia coli
2.3.1.46 1.8
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K47R Escherichia coli
2.3.1.46 2
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46R/K47R Escherichia coli
2.3.1.46 2
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46R/K47R Escherichia coli
2.3.1.46 4.1
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46A Escherichia coli
2.3.1.46 4.1
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46A Escherichia coli
2.3.1.46 37
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46R Escherichia coli
2.3.1.46 37
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46R Escherichia coli
2.3.1.46 130
-
 succinyl-CoA pH 7.2, 25°C, recombinant wild-type enzyme Escherichia coli
2.3.1.46 130
-
 L-homoserine pH 7.2, 25°C, recombinant wild-type enzyme Escherichia coli
2.3.1.46 130
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant C90S Escherichia coli
2.3.1.46 130
-
 L-homoserine pH 7.2, 25°C, recombinant mutant C90S Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.46 7.2
-
 assay at Escherichia coli

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.3.1.46 6.2 8.5 pH profile of mutant K47R Escherichia coli