Ceramide enables Fas to cap and kill Journal Article
| Authors: | Cremesti, A.; Paris, F.; Grassmé, H.; Holler, N.; Tschopp, J.; Fuks, Z.; Gulbins, E.; Kolesnick, R. |
| Article Title: | Ceramide enables Fas to cap and kill |
| Abstract: | Recent studies suggest that trimerization of Fas is insufficient for apoptosis induction and indicate that super-aggregation of trimerized Fas might be prerequisite. For many cell surface receptors, cross-linking by multivalent ligands or antibodies induces their lateral segregation within the plasma membrane and co-localization into "caps" on one pole of the cell. In this study, we show that capping of Fas is essential for optimal function and that capping is ceramide-dependent. In Jurkat T lymphocytes and in primary cultures of hepatocytes, ceramide elevation was detected as early as 15-30 s and peaked at 1 min after CH-11 and Jo2 anti-Fas antibody treatment, respectively. Capping was detected 30 s after Fas ligation, peaked at 2 min, and was maintained at a lower level for as long as 30 min in both cell types. Ceramide generation appeared essential for capping. Acid sphingomyelinase -/- hepatocytes were defective in Jo2-induced ceramide generation, capping, and apoptosis, and nanomolar concentrations of C16-ceramide restored these events. To further explore the role of ceramide in capping of Fas, we employed FLAG-tagged soluble Fas ligand (sFasL), which binds trimerized Fas but is unable to induce capping or apoptosis in Jurkat cells. Cross-linking of sFasL with M2 anti-FLAG antibody induced both events. Pretreatment of cells with natural C16-ceramide bypassed the necessity for forced antibody cross-linking and enabled sFasL to cap and kill. The presence of intact sphingolipid-enriched membrane domains may be essential for Fas capping since their disruption with cholesterol-depleting agents abrogated capping and prevented apoptosis. These data suggest that capping is a ceramide-dependent event required for optimal Fas signaling in some cells. |
| Keywords: | signal transduction; controlled study; unclassified drug; genetics; nonhuman; protein domain; t lymphocyte; t-lymphocytes; animal cell; mouse; animal; cytology; metabolism; mouse mutant; animals; mice; mice, knockout; cells, cultured; hepatocytes; apoptosis; fas antigen; fas ligand; protein binding; drug effect; physiology; cell type; animalia; biosynthesis; immunology; kinetics; cell culture; membrane glycoproteins; polymerization; membrane protein; cell membrane; ligand; cellular distribution; cholesterol; fas ligand protein; antibodies; liver cell; biochemistry; ligand binding; antibody; protein cross linking; concentration response; antibody production; receptor binding; leukemia cell line; jurkat cells; protein antibody; ceramide; ceramides; immunoglobulin m antibody; cell surface receptor; sphingomyelin phosphodiesterase; cell surface; ceramide derivative; cells; membrane structure; membrane microdomains; antigens, cd95; nystatin; cell disruption; sphingolipid; acid sphingomyelinase 1; acid sphingomyelinase-1; receptor aggregation; filipin; enzyme defect; humans; human; priority journal; article; capping phenomenon; crosslinking; trimerization; fasl protein, mouse; faslg protein, human; alpha cyclodextrin; beta cyclodextrin; c16 dihydroceramide; c6 ceramide; ch 11 antibody; flag antibody m2 |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 276 |
| Issue: | 26 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2001-06-29 |
| Start Page: | 23954 |
| End Page: | 23961 |
| Language: | English |
| DOI: | 10.1074/jbc.M101866200 |
| PUBMED: | 11287428 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 21 May 2015 -- Source: Scopus |
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