Separation and properties of two arylamidases from rat cardiac-muscle extracts.
Two main arylamidase activities were separated from a particle-free supernatant of rat heart by chromatography on DEAE-Sephadex. Although both enzymes hydrolysed L-leucine 4-nitroanilide, only peak-II enzyme hydrolysed L-lysine 4-nitroanilide. A third minor peak (Ia) contained an enzyme that was active mainly on the L-lysine 4-nitroanilide. The mol.wts. of the enzymes in peaks I and II were approx. 257000 and 105000 respectively. The pH optimum was approx. pH7.0 for peak-I enzyme and 7.0-8.0 for peak-II enzyme. Both enzymes were inhibited by addition of puromycin, p-hydroxymercuribenzoate, o-phenanthroline and bivalent metal ions. Addition of dithiothreitol resulted in stimulation of both activities. Dialysis against o-phenanthroline resulted in inhibition of peak-I and -II enzymes, but after dialysis against EDTA only peak-II enzyme was inhibited.
D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D000818:Animals; D002413:Cations, Divalent; D002852:Chromatography, Ion Exchange; D003956:Dialysis; D004229:Dithiothreitol / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D005260:Female; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D009206:Myocardium / Q000201:enzymology*; D010618:Phenanthrolines / Q000494:pharmacology; D011691:Puromycin / Q000494:pharmacology; D051381:Rats
D002413:Cations, Divalent; D006902:Hydroxymercuribenzoates; D010618:Phenanthrolines; D011691:Puromycin; D004492:Edetic Acid; D000626:Aminopeptidases; D004229:Dithiothreitol