Selective Inhibition of Dipeptidyl Peptidase I, Not Caspases, Prevents the Partial Processing of Procaspase-3 in CD3-activated Human CD8+ T Lymphocytes

• Published in: The Journal of biological chemistry Vol. 277; no. 35; pp. 32339 - 32347
• Main Authors: Nicolas Bidère, Marie Briet, Antoine Dürrbach, Céline Dumont, Jérôme Feldmann, Bernard Charpentier, Geneviève de Saint-Basile, Anna Senik
• Format: Journal Article
• Language: English
• Published: United States American Society for Biochemistry and Molecular Biology 30.08.2002
• Subjects: Amino Acid Chloromethyl Ketones - pharmacology; Antigens, CD - physiology; Caspase 3; Caspases - metabolism; Cathepsin C - antagonists & inhibitors; CD3 Complex - physiology; CD8-Positive T-Lymphocytes - immunology; Cysteine Proteinase Inhibitors - pharmacology; Enzyme Precursors - metabolism; Humans; Lymphocyte Activation; Protease Inhibitors - pharmacology; Protein Processing, Post-Translational; Protein Subunits; Protein Transport
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M205153200

Activation of primary human T cells by anti-CD3 and interleukin-2 resulted in partial processing of procaspase-3 in activated nonapoptotic (ΔΨ m high ) CD8 + T cells but not in CD4 + T cells. Apical caspases-8 and -9 were not activated, and Bid was not processed to truncated Bid. Boc-D.fmk, a broad spectrum caspase inhibitor, did not prevent this process, whereas GF.dmk, a selective inhibitor of dipeptidyl peptidase I, was effective. Dipeptidyl peptidase I is required for the activation of granule-associated serine proteases. It is enriched in the cytolytic granules of cytotoxic lymphocytes, where it promotes the proteolytic activation of progranzymes A and B. Inhibition of granzyme B (GrB)-like serine proteases by Z-AAD.cmk prevented partial processing of procapase-3, whereas inhibition of GrA activity by d -FPR.cmk had no effect. Specific inhibitors of other lysosomal proteases such as cathepsins B, L, and D did not interfere in this event. Patients with Chediak-Higashi syndrome or with perforin deficiency also displayed partial processing of procaspase-3, excluding the involvement of granule exocytosis for the delivery of the serine protease in cause. The p20/p12 processing pattern of procaspase-3 in our model points to GrB, the sole serine protease with aspase activity. Small amounts of GrB were indeed exported from cytolytic granules to the cytosol of a significant fraction of GrB-positive cells.