Inhibition by alpha-macroglobulin and other serum proteins.
1. Normal human serum was found to inhibit human cathepsin B1. 2. The major inhibitor present in serum was purified and identified as alpha(2)-macroglobulin. 3. alpha(2)-Macroglobulin was found to bind cathepsin B1 in an approximately 1:1 molar ratio. When bound, the enzyme retained about 50% of its proteolytic activity, and up to 80% of its activity against alpha-N-benzoyl-dl-arginine 2-naphthylamide. 4. Pretreatment of alpha(2)-macroglobulin with cathepsin B1 inactivated by exposure to pH8.5 or iodoacetic acid, in large molar excess, did not prevent the subsequent binding of active enzyme. Active enzyme, once bound, was not protected from inhibition by 1-chloro-4-phenyl-3-tosylamido-l-butan-2-one. 5. Cathepsin B1 was also inhibited by human immunoglobulin G, at high concentration. 6. Because it had been suggested that haptoglobin is responsible for the inhibition of ;cathepsin B' by serum, a method was devised for the selective removal of haptoglobin from mixtures of serum proteins by adsorption on haemoglobin covalently linked to Sepharose. No evidence was obtained that haptoglobin has any inhibitory activity against the enzyme.
D000818:Animals; D001168:Arthritis / Q000097:blood; D001665:Binding Sites; D001798:Blood Proteins; D002403:Cathepsins / Q000037:antagonists & inhibitors*; D002845:Chromatography; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G / Q000302:isolation & purification; D007461:Iodoacetates; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008259:Macroglobulins; D011134:Polysaccharides; D011485:Protein Binding; D011489:Protein Denaturation; D011817:Rabbits / Q000276:immunology; D013056:Spectrophotometry, Ultraviolet; D014462:Ultrafiltration
D001798:Blood Proteins; D007106:Immune Sera; D007074:Immunoglobulin G; D007461:Iodoacetates; D008259:Macroglobulins; D011134:Polysaccharides; D002403:Cathepsins