Release of a glycosylphosphatidylinositol-anchored ADP-ribosyltransferase from cytotoxic T cells upon activation.
Many cell surface proteins are anchored into the cell membrane by glycosylphosphatidylinositol (GPI), among those a recently discovered arginine-specific mono-ADP-ribosyltransferase on cytotoxic T cells (CTL). This enzyme transfers ADP-ribose to cell surface proteins resulting in inhibition of cytotoxic and proliferative activity. Here we report that ADP-ribosyltransferase is released in active forms by crosslinking CD3, exposure to Il-2 or PMA stimulation. Release of transferase is specific, as another GPI-anchored protein, Thy-1 is not released. Transferase molecules released by cell activation are indistinguishable in size from molecules released by phospholipase C, suggesting that the release mechanism acts close to or within the GPI anchor. Protease inhibitors fail to inhibit transferase release with exception of 1,10-phenanthroline and its 4,7-diphenyl derivative. This suggests that the release mechanism acts on the cell surface but does not discriminate between action of a metalloprotease or phospholipase D. Release of transferase is shown to be rapid, it is not suppressed by monensin or brefeldin A and independent of serum phospholipase D, consistent with a mechanism acting on the cell surface. Transferase expression is shown to be dependent on the cell activation stage. In CTL clones, the transferase is demonstrable as a phospholipase C releasable molecule at early but not later stages of Ag specific activation.
Nemoto
E
E
Department of Molecular Microbiology and Immunology, University of Southern California School of Medicine, Los Angeles 90033, USA.
Stohlman
S
S
Dennert
G
G
D036002:ADP Ribose Transferases; D000818:Animals; D004789:Enzyme Activation / Q000276:immunology; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D008213:Lymphocyte Activation; D051379:Mice; D008806:Mice, Inbred AKR; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D011065:Poly(ADP-ribose) Polymerases / Q000276:immunology / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic / Q000201:enzymology* / Q000378:metabolism*; D018800:Thy-1 Antigens / Q000096:biosynthesis; D014407:Tumor Cells, Cultured
D017261:Glycosylphosphatidylinositols; D018800:Thy-1 Antigens; D036002:ADP Ribose Transferases; D011065:Poly(ADP-ribose) Polymerases