title: Kinetic studies on the binding of gostatin, a suicide substrate for aspartate aminotransferase, with the isoenzymes from porcine heart mitochondria and cytosol.
abstract: The reaction of pig heart mitochondrial and cytosolic aspartate aminotransferases (abbreviated to mAspAT and cAspAT, respectively) with an enzyme-suicide substrate (mechanism-based inhibitor), gostatin (5-amino-2-carboxyl-4-oxo-1,4,5,6-tetrahydropyridine-3-acetic acid) was studied kinetically, by following the spectral change with a micro-stopped-flow apparatus, as well as the inactivation of the enzyme activity. No significant difference in kinetic behavior was observed between mAspAT and cAspAT. From the analysis of time-dependent spectral change, no positive evidence for the existence of spectrophotometrically distinguishable intermediates was obtained. Both the spectral change and the inactivation followed, at least in appearance, simple bimolecular association kinetics, under the conditions studied. However, the second-order rate constant of the spectral change was found to be 1.5 to 2 times as large as that of the inactivation. The effects of pH and temperature on k(on) (the second-order rate constant of the spectral change) were also studied.
authors.affiliation: Faculty of Agriculture, Kyoto University.
authors.forename: K
authors.initials: K
authors.lastname: Kitagishi
authors.forename: K
authors.initials: K
authors.lastname: Hiromi
authors.forename: S
authors.initials: S
authors.lastname: Tanase
authors.forename: F
authors.initials: F
authors.lastname: Nagashima
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
mesh_terms: D000600:Amino Acids, Dicarboxylic / Q000378:metabolism*; D000818:Animals; D001219:Aspartate Aminotransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D003600:Cytosol / Q000201:enzymology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007700:Kinetics; D008929:Mitochondria, Heart / Q000201:enzymology*; D009206:Myocardium / Q000378:metabolism*; D011485:Protein Binding; D013053:Spectrophotometry; D013552:Swine
chemical_list: D000600:Amino Acids, Dicarboxylic; D007527:Isoenzymes; C042220:gostatin; D001219:Aspartate Aminotransferases