A rapid and simple method for purification of the chloride-activated arginine aminopeptidase from biological materials.
A homogenous, chloride-dependent arginine amino-peptidase was purified from the liver of human fetuses by gel-permeation chromatography followed by subsequent fractionation on DEAE-Sepharose CL-6B and affinity chromatography on Sepharose 4B covalently coupled to L-arginine. The purified enzyme showed a single band on disc-gel electrophoresis. In SDS-gel electrophoresis the molecular weight of the enzyme was found to be 92'000 +/- 2000. N-L-Arginyl-2-naphthylamine and N-L-lysyl-2-naphthylamine were practically the only aminoacyl-2-naphthylamines hydrolyzed by the enzyme. The method was successfully applied for the purification of the chloride-dependent arginine aminopeptidases from human erythrocytes, serum, synovial fluid and rat inflammatory exudates.
D000626:Aminopeptidases / Q000097:blood / Q000302:isolation & purification*; D000818:Animals; D001120:Arginine / Q000097:blood / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D004589:Electrophoresis, Disc; D004912:Erythrocytes / Q000201:enzymology; D005122:Exudates and Transudates / Q000201:enzymology; D005333:Fetus / Q000201:enzymology*; D006801:Humans; D008099:Liver / Q000201:enzymology*; D051381:Rats; D013582:Synovial Fluid / Q000201:enzymology
D001120:Arginine; D000626:Aminopeptidases; C019721:aminopeptidase B