title: Isolation and characterization of a new aminopeptidase from bovine lens.
abstract: An aminopeptidase has been purified to homogeneity from bovine lens tissue by gel filtration and DEAE-cellulose chromatography. This enzyme has a molecular weight of 96,000 under both native and denaturing conditions. The purified enzyme hydrolyzed a variety of synthetic substrates as well as di-, tri-, and higher molecular weight peptides. Significantly this enzyme is capable of hydrolyzing arginine, lysine, and proline aminoacyl bonds. The pH optimum for activity and stability was 6.0. Both a reduced sulfhydryl group and a divalent metal ion are essential for activity. The native enzyme contains 1.6 mol of zinc and 1.0 mol of copper/mol of enzyme. No activation was seen upon incubation with either magnesium or manganese; however, heavy metal ions were inhibitory. Bestatin and puromycin were effective inhibitors and no endopeptidase activity could be detected in the purified preparation. This enzyme is clearly distinct from the lens leucine aminopeptidase, but rather, is identical to a cytosolic aminopeptidase III isolated from other tissues. Evidence is presented which argues that this enzyme may be the major lens aminopeptidase under in vivo conditions.
authors.forename: K K
authors.initials: KK
authors.lastname: Sharma
authors.forename: B J
authors.initials: BJ
authors.lastname: Ortwerth
mesh_terms: D000626:Aminopeptidases / Q000302:isolation & purification*; D000818:Animals; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003300:Copper / Q000032:analysis; D004492:Edetic Acid / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007908:Lens, Crystalline / Q000201:enzymology*; D008970:Molecular Weight; D029701:Saccharomyces cerevisiae Proteins; D013696:Temperature; D015032:Zinc / Q000032:analysis
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D003300:Copper; D004492:Edetic Acid; D000626:Aminopeptidases; C019689:APE1 protein, S cerevisiae; D015032:Zinc