Purification and properties of rabbit liver cathepsin M and cathepsin B.
Cathepsins M and B from rabbit liver lysosomes were separated by chromatography on Ultrogel AcA34 at low ionic strength and purified to homogeneity, and their catalytic and molecular properties were compared. Cathepsin M was relatively inactive with synthetic peptide substrates. Thus, it hydrolyzed benzoyl arginine naphthylamide at only one-fifth the rate observed with cathepsin B, and no activity was detected with Gly-Phe naphthylamide which is a relatively good substrate for cathepsin B. On the other hand, cathepsin M exhibited a preference for protein substrates. It was more active than cathepsin B in catalyzing the inactivation of the following enzymes: rabbit muscle or liver fructose-1,6-bisphosphate aldolases, rabbit liver fructose-1,6-bisphosphatase and pyruvate kinase, yeast glucose-6-phosphate dehydrogenase, and rabbit muscle glyceraldehyde-3-phosphate dehydrogenase. With glucagon as substrate, both enzymes showed similar peptidyl dipeptidase activities with some minor differences in peptide bond specificity. Cathepsins M and B are similar in size, with apparent molecular weights of 30,200 for cathepsin M and 28,800 for cathepsin B, and in amino acid composition and carbohydrate content. Each contains approximately 2-3 equivalents/mol glucosamine, 3 equivalents/mol mannose, and no fucose or galactosamine. They also show similar microheterogeneity in sodium dodecylsulfate-gel electrophoresis and isoelectric focusing; this microheterogeneity is probably related to differences in glycosylation. Extensive homology in primary structure for the two proteins was indicated by the similar patterns of peptides formed on digestion with trypsin.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001587:Benzoylarginine-2-Naphthylamide / Q000378:metabolism; D002241:Carbohydrates / Q000032:analysis; D002401:Cathepsin B; D002403:Cathepsins / Q000302:isolation & purification* / Q000378:metabolism; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D005934:Glucagon / Q000494:pharmacology; D006596:Hexosaminidases / Q000378:metabolism; D007525:Isoelectric Focusing; D008099:Liver / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D009281:Naphthalenes / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis; D011817:Rabbits; D013379:Substrate Specificity; D014357:Trypsin / Q000378:metabolism
D000596:Amino Acids; D002241:Carbohydrates; D009281:Naphthalenes; D010446:Peptide Fragments; D001587:Benzoylarginine-2-Naphthylamide; D005934:Glucagon; D006596:Hexosaminidases; D002403:Cathepsins; D014357:Trypsin; D002401:Cathepsin B; C034114:cathepsin M