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Literature summary extracted from

  • Ozaki, S.; Yoshiya, M.
    Mutagenesis of arginine-186 located on a helix interacting with pyridoxal 5'-phosphate in O-acetylserine sulfhydrylase (2008), Asian J. Biochem., 3, 351-358.
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.47 expression of the glutathione S-transferase (GST)-fusion enzyme in Escherichia coli BL21 with pGEX 4T-2 vector Salmonella enterica subsp. enterica serovar Typhimurium

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.47 R186L retains one cofactor per subunit, accelerates the reaction with substrate O3-acetyl-L-serine 1.8fold, intermediates are formed faster by 1.5 and 1.3fold, respectively, with azide or thiosulfate than in the wild type Salmonella enterica subsp. enterica serovar Typhimurium
2.5.1.47 R186P loss of cofactor leads to enzyme inactivation Salmonella enterica subsp. enterica serovar Typhimurium

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.5.1.47 35000
-
 SDS-PAGE 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
-
 ?

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.47 cell harvesting are resuspended in 50 mM Tris buffer, pH 8.0, containing 10 mM EDTA, 15 mM beta-mercaptoethanol, 0.05 mM N-p-tosyl-L-lysine chloromethylketone, leupeptin, aprotinin, pepstatin, lysozyme and pyridoxal 5'-phosphate, centrifuged, supernatant loaded on a DE52 anion exchange column, washed with 50 mM Tris buffer, pH 8.0, with 10 mM EDTA, and 15 mM beta-mercaptoethanol, elution with NaCl gradient in buffer, active fractions are pooled, concentrated and applied to a GSTrap 4B column, washed with PBS buffer, pH 7.4, containing 140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, and 1.8 mM KH2PO4, elution with 50 mM Tris, pH 8.0, with 20 mM reduced glutathione and thrombin, dialyzed with 15 mM potassium phosphate, pH 7.2, applied on a DE 52 column, separation of enzyme from GST-tag and thrombin with a gradient of 15-300 mM potassium phosphate buffer, pH 7.2 Salmonella enterica subsp. enterica serovar Typhimurium

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.47 O3-acetyl-L-serine + hydrogen sulfide
-
 Salmonella enterica subsp. enterica serovar Typhimurium 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

Cofactor

EC Number Cofactor Comment Organism Structure
2.5.1.47 pyridoxal 5'-phosphate R186 interacts with the cofactor Salmonella enterica subsp. enterica serovar Typhimurium