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Literature summary extracted from

  • Tai, C.H.; Rabeh, W.M.; Guan, R.; Schnackerz, K.D.; Cook, P.F.
    Role of histidine-152 in cofactor orientation in the PLP-dependent O-acetylserine sulfhydrylase reaction (2008), Arch. Biochem. Biophys., 472, 115-125.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.47 expression in with plasmid pRSM40 in Escherichia coli NM522 Salmonella enterica subsp. enterica serovar Typhimurium

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.47 H152A shift in the ketoenamine to enolimine tautomeric equilibrium toward the neutral enolimineine, the internal Schiff base of the free enzyme, the amino acid external Schiff base, and the alpha-aminoacrylate intermediate. The decreased rate of the mutant likely reflects a decrease in the amount of active enzyme as a result of an increased flexibility of the cofactor , which leads to increased rates of interconversion of the open and closed forms of the enzyme and additional interactions between the cofactor and enzyme in the closed form of the enzyme. Analysis of spectral properties Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 H152A shift in the ketoeneamine to enolimine tautomeric equilibrium towards neutral enolimine in the internal Shiff base of the free enzyme, the amino acid external Schiff base, and the alpha-aminoacrylate intermediate, 2 enzyme conformers are present, decreased rate of the enzyme likely reflects a decrease in the amount of active enzyme as result of the increased cofactor flexibility which stabilizes the nonproductive binding of O3-acetyl-L-serine in the external Schiff base Salmonella enterica subsp. enterica serovar Typhimurium

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.47 0.11
-
O-acetyl-L-serine mutant H52A, pH 6.5 Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 0.11
-
O3-acetyl-L-serine H152A mutant with 136fold decrease compared to wild type, 100 mM MES, pH 6.5, 50 microM 5-thio-2-nitrobenzoate (TNB), 25°C Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 0.6
-
5-thio-2-nitrobenzoate wild-type, pH 7.0 Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 0.6
-
5-thio-2-nitrobenzoate wild type, 100 mM HEPES, pH 7.0, 0.5 mM O3-acetyl-L-serine, 25°C Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 2.5
-
5-thio-2-nitrobenzoate mutant H52A, pH 6.5 Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 2.5
-
5-thio-2-nitrobenzoate H152A mutant with 4.1fold increase compared to wild type, 100 mM MES, pH 6.5, 0.5 mM O3-acetyl-L-serine, 25°C Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 15
-
O-acetyl-L-serine wild-type, pH 7.0 Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 15
-
O3-acetyl-L-serine wild type, 100 mM HEPES, pH 7.0, 50 microM 5-thio-2-nitrobenzoate (TNB), 25°C Salmonella enterica subsp. enterica serovar Typhimurium

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.47 O3-acetyl-L-serine + hydrogen sulfide Salmonella enterica subsp. enterica serovar Typhimurium
-
L-cysteine + acetate
-
?
2.5.1.47 O3-acetyl-L-serine + hydrogen sulfide Salmonella enterica subsp. enterica serovar Typhimurium DW378
-
L-cysteine + acetate
-
?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.47 Salmonella enterica subsp. enterica serovar Typhimurium
-
-
-
2.5.1.47 Salmonella enterica subsp. enterica serovar Typhimurium
-
DW378
-
2.5.1.47 Salmonella enterica subsp. enterica serovar Typhimurium DW378
-
DW378
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.47 O-acetyl-L-serine + 5-thio-2-nitrobenzoate
-
Salmonella enterica subsp. enterica serovar Typhimurium ? + acetate
-
?
2.5.1.47 O-acetyl-L-serine + 5-thio-2-nitrobenzoate
-
Salmonella enterica subsp. enterica serovar Typhimurium DW378 ? + acetate
-
?
2.5.1.47 O3-acetyl-L-serine + 5-thio-2-nitrobenzoate
-
Salmonella enterica subsp. enterica serovar Typhimurium ? + acetate
-
?
2.5.1.47 O3-acetyl-L-serine + 5-thio-2-nitrobenzoate
-
Salmonella enterica subsp. enterica serovar Typhimurium DW378 ? + acetate
-
?
2.5.1.47 O3-acetyl-L-serine + hydrogen sulfide
-
Salmonella enterica subsp. enterica serovar Typhimurium L-cysteine + acetate
-
?
2.5.1.47 O3-acetyl-L-serine + hydrogen sulfide
-
Salmonella enterica subsp. enterica serovar Typhimurium DW378 L-cysteine + acetate
-
?

Synonyms

EC Number Synonyms Comment Organism
2.5.1.47 O-acetylserine sulfhydrylase
-
Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 OASS-A
-
Salmonella enterica subsp. enterica serovar Typhimurium

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.5.1.47 additional information
-
additional information wild type and mutant exhibit almost identical values for sulfide as substrate, imidazole has no effect on the activity of wild type or H152A mutant enzyme Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 0.06
-
O3-acetyl-L-serine the turnover for H152A mutant is decreased 9fold compared to the wild type, 25°C, 100 mM MES, pH 6.5 Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 0.56
-
O3-acetyl-L-serine 25°C, 100 mM HEPES, pH 7.0 Salmonella enterica subsp. enterica serovar Typhimurium

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.5.1.47 additional information
-
pH-rate profiles for wild type and H152A mutant with O3-acetyl-L-serine and 5-thio-2-nitrobenzoate as substrates are similar, 100 mM concentrations of 2-(N-morpholino)ethanesulfonic acid (MES), pH 5.5-6.5, 3-(N-morpholino)propanesulfonic acid (MOPS), pH 7.0-7.5, N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid (HEPES), pH 8.0, and 3-[[tris(hydroxymethyl)]amino]propanesulfonic acid (TAPS), pH 9.0, titrated with KOH Salmonella enterica subsp. enterica serovar Typhimurium

Cofactor

EC Number Cofactor Comment Organism Structure
2.5.1.47 pyridoxal 5'-phosphate the 5'-phosphate is tightly bound to the enzyme via a hydrogen bond to His152, one to the residues responsible for positioning of the cofactor Salmonella enterica subsp. enterica serovar Typhimurium

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.5.1.47 additional information
-
additional information wild type and mutant exhibit almost identical values for sulfide as substrate Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 0.0219
-
5-thio-2-nitrobenzoate H152A mutant with 43fold decrease compared to wild type, 100 mM MES, pH 6.5, 0.5 mM O3-acetyl-L-serine, 25°C Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 0.037
-
O3-acetyl-L-serine wild type, 100 mM HEPES, pH 7.0, 50 microM 5-thio-2-nitrobenzoate (TNB), 25°C Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 0.51
-
O3-acetyl-L-serine H152A mutant with 14fold increase compared to wild type, 100 mM MES, pH 6.5, 50 microM 5-thio-2-nitrobenzoate (TNB), 25°C Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 0.95
-
5-thio-2-nitrobenzoate wild type, 100 mM HEPES, pH 7.0, 0.5 mM O3-acetyl-L-serine, 25°C Salmonella enterica subsp. enterica serovar Typhimurium