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Literature summary for 2.5.1.47 extracted from

  • Tai, C.H.; Rabeh, W.M.; Guan, R.; Schnackerz, K.D.; Cook, P.F.
    Effect of mutation of lysine-120, located at the entry to the active site of O-acetylserine sulfhydrylase-A from Salmonella typhimurium (2008), Biochim. Biophys. Acta, 1784, 629-637.
    View publication on PubMed
Search for more literature for 2.5.1.47

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli NM522 with plasmid pRSM40 Salmonella enterica subsp. enterica serovar Typhimurium

Protein Variants

Protein Variants Comment Organism
K120Q mutation results in a shift in the tautomeric equilibrium toward the neutral enolimine and an increase in the rate of interconversion of the open and closed forms of the enzyme. A decrease in the rate of both half reactions reflects the stabilization of the external Schiff base. Role of K120 in helping to stabilize the closed conformation of the enzyme by participating in a new bond to the backbone carbonyl of A231 Salmonella enterica subsp. enterica serovar Typhimurium
K120Q shift in the tautomeric equilibrium toward the neutral enolimine and an increase of the rate of interconversion of the open and closed forms of the enzyme, probably reflecting the stabilization of the external Schiff base Salmonella enterica subsp. enterica serovar Typhimurium

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.07
-
 5-thio-2-nitrobenzoate mutant H52A, pH 6.5 Salmonella enterica subsp. enterica serovar Typhimurium
0.07
-
 5-thio-2-nitrobenzoate K120Q mutant with 9fold 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
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
1
-
 O-acetyl-L-serine mutant H52A, pH 6.5 Salmonella enterica subsp. enterica serovar Typhimurium
1
-
 O3-acetyl-L-serine K120Q mutant with 15fold 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
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)

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

Organism

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

Purification (Commentary)

Purification (Comment) Organism
Q-Sepharose and phenyl Sepharose column, active fractions are pooled and concentrated Salmonella enterica subsp. enterica serovar Typhimurium

Substrates and Products (Substrate)

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

Synonyms

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
 O3-acetyl-L-serine turnover for mutant K120Q is decreased 56fold compared to wild type, 25°C, 100 mM HEPES, pH 7.0 Salmonella enterica subsp. enterica serovar Typhimurium
0.56
-
 O3-acetyl-L-serine wild type, 25°C, 100 mM HEPES, pH 7.0 Salmonella enterica subsp. enterica serovar Typhimurium

pH Range

pH Minimum pH Maximum Comment Organism
additional information
-
 pH-rate profiles for wild type and H152A mutant with O3-acetyl-L-serine and beta-chloro-L-alanine 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

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate
-
 Salmonella enterica subsp. enterica serovar Typhimurium

General Information

General Information Comment Organism
metabolism catalyzes the final step of the L-cysteine biosynthesis Salmonella enterica subsp. enterica serovar Typhimurium

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.01
-
 O3-acetyl-L-serine K120Q mutant with 3.7fold decrease compared to wild type value, 100 mM MES, pH 6.5, 50 microM 5-thio-2-nitrobenzoate (TNB), 25°C Salmonella enterica subsp. enterica serovar Typhimurium
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
0.141
-
 5-thio-2-nitrobenzoate K120Q mutant with 6.7fold decrease compared to wild type value, 100 mM MES, pH 6.5, 0.5 mM O3-acetyl-L-serine, 25°C Salmonella enterica subsp. enterica serovar Typhimurium
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