Caspase activity is required for commitment to Fas-mediated apoptosis.
Recognition of the widespread importance of apoptosis has been one of the most significant changes in the biomedical sciences in the past decade. The molecular processes controlling and executing cell death through apoptosis are, however, still poorly understood. The ICE (Interleukin-1beta Converting Enzyme) family-recently named the caspases for cysteine aspartate-specific proteases-plays a central role in apoptosis and may well constitute part of the conserved core mechanism of the process. Potentially, these proteases may be of great significance, both in the pathology associated with failure of apoptosis and also as targets for therapeutic intervention where apoptosis occurs inappropriately, e.g. in degenerative disease and AIDS. However, this is only likely if caspase activity is required before commitment to mammalian cell death. Here, we have used both peptide inhibitors and crmA transfection to inhibit these proteases in intact cells. Our experiments show that selective inhibition of some caspases protects human T cells (Jurkat and CEM-C7) from Fas-induced apoptosis, dramatically increasing their survival (up to 320-fold) in a colony-forming assay. This suggests that dysfunction of some, but not all, caspases could indeed play a crucial part in the development of some cancers and autoimmune disease, and also that these proteases could be appropriate molecular targets for preventing apoptosis in degenerative disease.
Longthorne
V L
VL
Department of Biological Sciences, Keele University, Staffordshire, UK.
Williams
G T
GT
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D017209:Apoptosis; D002455:Cell Division; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D006801:Humans; D019169:Jurkat Cells; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D015843:Serpins / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000201:enzymology; D014407:Tumor Cells, Cultured; D014764:Viral Proteins; D019014:fas Receptor / Q000378:metabolism*
D000590:Amino Acid Chloromethyl Ketones; D015853:Cysteine Proteinase Inhibitors; C098738:N-acetyl-tyrosyl-valyl-alanyl-aspartyl chloromethyl ketone; D015843:Serpins; D014764:Viral Proteins; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D019014:fas Receptor; C074641:interleukin-1beta-converting enzyme inhibitor; D011065:Poly(ADP-ribose) Polymerases; D003546:Cysteine Endopeptidases