title: Acetylation of Lys-373 in porcine pancreatic lipase after reaction of the enzyme or its C-terminal fragment [corrected] with p-nitrophenyl acetate.
abstract: The reactions of lipase (449 amino acid residues) and lipase fragment (336-449) with p-nitrophenyl acetate have been studied from 2 different angles. In previous papers it has been shown that lipase and lipase fragment enzymatically hydrolyze p-nitrophenyl acetate. The amino acid residue of the catalytic site that is temporarily acetylated has not yet been characterized in lipase or lipase fragment. Besides this very fast enzymatic hydrolysis, acetylation reactions may take place on nucleophilic amino acid side-chain groups. In the present report, acetylated amino acid residues whose acetyl linkages were not cleaved after pH 7.5-8.5 incubations have been investigated. Several residues were acetylated in very low proportion, whereas lysine 373 was stoichiometrically acetylated in lipase and in lipase fragment. This specific acetylation may have been favored by the presence of a hydrophobic reversible binding site for p-nitrophenyl acetate near Lys-373. This acetylation did not greatly change the specific activity of lipase towards an emulsion of tributyrylglycerol in the presence of colipase, but under certain conditions it had an effect on the enzymatic hydrolysis of p-nitrophenyl acetate by the lipase fragment.
authors.affiliation: Centre de Biochimie et de Biologie Moléculaire du Centre National de la Recherche Scientifique, Marseille, France.
authors.forename: J D
authors.initials: JD
authors.lastname: De Caro
authors.forename: M P
authors.initials: MP
authors.lastname: Chautan
authors.forename: P
authors.initials: P
authors.lastname: Rouimi
authors.forename: M
authors.initials: M
authors.lastname: Rovery
mesh_terms: D000107:Acetylation; D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008049:Lipase; D008239:Lysine; D009596:Nitrophenols; D010179:Pancreas / Q000201:enzymology*; D011817:Rabbits; D013552:Swine
chemical_list: D009596:Nitrophenols; C008642:4-nitrophenyl acetate; D008049:Lipase; D008239:Lysine