The nature of the collagenolytic cathepsin of rat liver and its distribution in other rat tissues.
1. An enzyme present in rat liver extracts degraded insoluble collagen maximally at pH3.5. Collagenolytic activity was more abundant in kidney, spleen and bone marrow and was also present in decreasing concentrations in ileum, lung, heart, skin and muscle. 2. The crude collagenolytic cathepsin was activated by cysteine and dithiothreitol, but not by 2-mercaptoethanol. Iodoacetamide, p-chloromercuribenzoate and 7-amino-1-chloro-3-l-tosylamidoheptan-2-one hydrochloride inhibited the enzyme. Zn(2+), Fe(3+) and Hg(2+) ions were strongly inhibitory, but Ca(2+), Co(2+), Mg(2+) and Fe(2+) ions had little or no effect. EDTA was an activator of the enzyme. Inhibitors of cathepsin B were found to enhance collagenolysis, but phenylpyruvic acid, a cathepsin D inhibitor, inhibited the enzyme. Di-isopropyl phosphorofluoridate had no effect. 3. Collagenolysis at pH3.5 and 28 degrees C was restricted to cleavage of the telopeptide region in insoluble collagen, and the material that was solubilized consisted mostly of alpha-chains. 4. The collagenolytic cathepsin was separated from cathepsins B2 and D by fractionation on Sephadex G-100 and a partial separation from cathepsin B1 was obtained by chromatography on DEAE-Sephadex. 5. The function of the collagenolytic cathepsin in the catabolism of collagen is discussed in relation to the action of the other lysosomal proteinases and the neutral collagenase.
D000818:Animals; D001853:Bone Marrow / Q000201:enzymology; D002118:Calcium; D002403:Cathepsins / Q000302:isolation & purification*; D002845:Chromatography; D003094:Collagen; D003545:Cysteine; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007461:Iodoacetates; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology*; D010667:Phenylpyruvic Acids; D011134:Polysaccharides; D051381:Rats; D013154:Spleen / Q000201:enzymology; D015032:Zinc
D007461:Iodoacetates; D010667:Phenylpyruvic Acids; D011134:Polysaccharides; D003094:Collagen; D002403:Cathepsins; D015032:Zinc; D003545:Cysteine; D002118:Calcium