title: Determination of human aspartate aminotransferase isoenzymes by their differential sensitivity to proteases.
abstract: The effect of various proteases (trypsin, chymotrypsin, subtilisin, protease 401, and thermolysin) on the mitochondrial isoenzyme (m-AST) and cytoplasmic isoenzyme (c-AST) of human and swine aspartate aminotransferase (AST;EC 2.6.1.1) was evaluated. All procedures including the reaction with proteases and the subsequent determination of the AST activity were carried out in an automatic analyzer. The mammalian c-AST was efficiently inactivated by chymotrypsin, subtilisin and protease 401 while m-AST activity decreased very slowly with these proteases. Thermolysin and trypsin showed much less effect on c-AST activity. Especially, chymotrypsin at concentrations of 0.5-1.0 g/L inactivated human c-AST almost completely but showed no detectable inactivating effect on m-AST. Thus chymotrypsin appears to be the most suitable protease for the differential determination of AST isoenzymes in human serum. Further studies on the effects of proteases with AST from other species showed that Escherichia coli AST resembled mammalian m-AST while Pseudomonas AST resembled c-AST.
authors.affiliation: Eiken Chemical Company, Tokyo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Teranishi
authors.forename: Y
authors.initials: Y
authors.lastname: Morino
authors.forename: T
authors.initials: T
authors.lastname: Nishino
authors.forename: S
authors.initials: S
authors.lastname: Murao
authors.forename: T
authors.initials: T
authors.lastname: Yagi
authors.forename: H
authors.initials: H
authors.lastname: Kagamiyama
authors.forename: N
authors.initials: N
authors.lastname: Harada
mesh_terms: D000818:Animals; D001219:Aspartate Aminotransferases / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D006801:Humans; D006868:Hydrolysis; D007527:Isoenzymes / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism; D009206:Myocardium / Q000201:enzymology; D010447:Peptide Hydrolases / Q000378:metabolism*; D011549:Pseudomonas / Q000201:enzymology; D013045:Species Specificity; D013379:Substrate Specificity; D013552:Swine
chemical_list: D007527:Isoenzymes; D001219:Aspartate Aminotransferases; D010447:Peptide Hydrolases