Lysosomal cysteine endopeptidases mediate interleukin 1-stimulated cartilage proteoglycan degradation.
The peptidyl diazomethane inactivator of cysteine endopeptidases, benzyloxycarbonyl-Tyr-Ala-CHN2, was tested as an inhibitor of interleukin 1 alpha-stimulated release of proteoglycan from bovine nasal septum cartilage explants. Like the previously tested epoxidyl peptide proinhibitor trans-epoxysuccinyl-leucylamido-(3-methyl)butane ethyl ester, it proved to be an effective inhibitor of proteoglycan release from cartilage, with significant inhibition at a concentration of 1 microM. The inhibition did not seem to be due to a general toxic effect. The rates of inactivation of the bovine cysteine endopeptidases by the peptidyl diazomethane, the epoxidyl peptide proinhibitor and its active form were determined. Benzyloxycarbonyl-Tyr-Ala-CHN2 proved to be a rapid inactivator of cathepsins L, S and B, but reacted much more slowly with cathepsin H and calpain. Thus it would appear that the latter two enzymes are not implicated in proteoglycan release in our test system. The peptidyl diazomethane and epoxidyl peptide proinhibitor (above) were also tested for their effects on three other interleukin 1-mediated cellular events, namely epidermal growth factor receptor transmodulation, and interleukin 6 and prostaglandin E2 production. In all cases the inactivators did not interfere with the response to interleukin 1 in human gingival fibroblasts. We conclude that one or more of the lysosomal cysteine endopeptidases cathepsins B, L and S mediate interleukin 1-stimulated cartilage proteoglycan degradation without affecting signal transduction.
Buttle
D J
DJ
Department of Biochemistry, Strangeways Research Laboratory, Worts Causeway, Cambridge, U.K.
Saklatvala
J
J
D000818:Animals; D002356:Cartilage / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D002401:Cathepsin B / Q000037:antagonists & inhibitors; D002417:Cattle; D046508:Culture Techniques; D003546:Cysteine Endopeptidases / Q000187:drug effects / Q000502:physiology*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology*; D003978:Diazomethane / Q000031:analogs & derivatives / Q000494:pharmacology; D004151:Dipeptides / Q000494:pharmacology; D004789:Enzyme Activation; D007375:Interleukin-1 / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D007700:Kinetics; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology*; D011509:Proteoglycans / Q000378:metabolism*; D013268:Stimulation, Chemical
D015853:Cysteine Proteinase Inhibitors; D004151:Dipeptides; D007375:Interleukin-1; D011509:Proteoglycans; C057752:benzyloxycarbonyltyrosylalanine diazomethane; D003978:Diazomethane; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; D007930:Leucine; C108192:aloxistatin