Generation of biologically active, complement-(C5) derived peptides by cathepsin H.
The thiol proteinase cathepsin H, isolated and purified from rat liver lysosomes, provokes acute inflammation characterized by the accumulation of polymorphonuclear leukocytes (PMN) when injected intracutaneously into newborn rats. We have examined the possibility that the accumulation of PMN at skin sites injected with cathepsin H is due, in part, to generation locally of C-derived chemotactic factors. We have found that cathepsin H acts in a concentration- and time-dependent fashion in whole human (and rat) EDTA-plasma to generate C5-derived peptides with chemotactic activity for PMN. Chemotactic activity was not generated in EDTA-plasma by either heat-inactivated cathepsin H or by a combination of active enzyme and a thiol proteinase inhibitor isolated from rat epidermis. Cathepsin H also acted in a concentration- and time-dependent fashion on isolated (functionally pure) human C5 to yield chemotactic activity for PMN as well as PMN lysosomal enzyme-releasing activity. Whereas 10 ng/ml cathepsin H generated significant chemotactic activity from isolated C5 (1000 CH50 U/ml), 7 to 10 micrograms/ml were required to generate chemotactic activity in whole EDTA-plasma. Cathepsin H not only was capable of generating biologically active, C5-derived peptides, but also was capable of degrading these peptides. Incubation of either whole EDTA-plasma or isolated C5 with high concentrations of cathepsin H (e.g., 25 micrograms/ml and 100 ng/ml, respectively) caused the rapid appearance of chemotactic activity followed by an equally rapid disappearance. PMN accumulated more rapidly in the skin of newborn rats injected with cathepsin H-treated C5 than in the skin of animals injected with cathepsin H alone. These data suggest that generation by cathepsin H of C-derived chemotactic activity contributes to the ability of this enzyme to induce dermal inflammation.
D000818:Animals; D056655:Cathepsin H; D002403:Cathepsins / Q000097:blood / Q000502:physiology*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D003182:Complement C5 / Q000008:administration & dosage / Q000378:metabolism*; D003546:Cysteine Endopeptidases; D003877:Dermatitis, Contact / Q000209:etiology / Q000473:pathology; D004492:Edetic Acid / Q000494:pharmacology; D006801:Humans; D008247:Lysosomes / Q000201:enzymology; D009504:Neutrophils / Q000201:enzymology; D010446:Peptide Fragments / Q000096:biosynthesis*; D011480:Protease Inhibitors / Q000097:blood; D051381:Rats; D011919:Rats, Inbred Strains
D003182:Complement C5; D010446:Peptide Fragments; D011480:Protease Inhibitors; D004492:Edetic Acid; D002403:Cathepsins; D003546:Cysteine Endopeptidases; C534126:CTSH protein, human; D056655:Cathepsin H; C534128:Ctsh protein, rat