title: Arylamine N-acetyltransferase activity in Staphylococcus aureus.
abstract: N-Acetyltransferase (NAT) activities were determined by incubation of Staphylococcus aureus cytosols with p-aminobenzoic acid (PABA) or 2-aminofluorene (2-AF) followed by high pressure liquid chromatography assays. The NAT activities from S. aureus were found to be 0.67 +/- 0.04 nmol/min/mg protein for the acetylation of 2-AF and 0.46 +/- 0.02 nmol/min/mg protein for the acetylation of PABA. The apparent K(m) and Vmax values obtained were 2.85 +/- 0.65 mM and 7.51 +/- 0.86 nmol/min/mg protein for 2-AF, and 2.35 +/- 0.39 mM and 9.43 +/- 0.78 nmol/min/mg protein for PABA, respectively. The optimal pH value for the enzyme activity was 7.0 for both substrates tested. The optimal temperature for enzyme activity was 37 degrees C for both substrates. The NAT activity was inhibited by iodoacetamide at 0.25 mM, and activity was reduced 50%. At 1.0 mM iodoacetamide activity was inhibited more than 90%. Among a series of divalent cations and salts, Zn2+, Ca2+, and Fe2+ were demonstrated to be the most potent inhibitors. The molecular weight of NAT from S. aureus was found to be 44.9 kDa. This report is the first demonstration of acetyl CoA: arylamine NAT activity in S. aureus.
authors.affiliation: Department of Medicine, China Medical College, Taichung, Taiwan, Republic of China.
authors.forename: F C
authors.initials: FC
authors.lastname: Chang
authors.forename: J G
authors.initials: JG
authors.lastname: Chung
authors.forename: W C
authors.initials: WC
authors.lastname: Chang
authors.forename: L T
authors.initials: LT
authors.lastname: Wu
authors.forename: G W
authors.initials: GW
authors.lastname: Chen
authors.forename: S H
authors.initials: SH
authors.lastname: Chang
mesh_terms: D001191:Arylamine N-Acetyltransferase / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007460:Iodoacetamide / Q000494:pharmacology; D007700:Kinetics; D013211:Staphylococcus aureus / Q000201:enzymology*; D013696:Temperature
chemical_list: D001191:Arylamine N-Acetyltransferase; D007460:Iodoacetamide