| First Author | Schneider P | Year | 1997 |
| Journal | J Biol Chem | Volume | 272 |
| Issue | 30 | Pages | 18827-33 |
| PubMed ID | 9228058 | Mgi Jnum | J:41925 |
| Mgi Id | MGI:894837 | Doi | 10.1074/jbc.272.30.18827 |
| Citation | Schneider P, et al. (1997) Characterization of Fas (Apo-1, CD95)-Fas ligand interaction. J Biol Chem 272(30):18827-33 |
| abstractText | The death-inducing receptor Fas is activated when cross-linked by the type II membrane protein Fas ligand (FasL). When human soluble FasL (sFasL, containing the extracellular portion) was expressed in human embryo kidney 293 cells, the three N-linked glycans of each FasL monomer were found to be essential for efficient secretion. Based on the structure of the closely related lymphotoxin alpha-tumor necrosis factor receptor I complex, a molecular model of the FasL homotrimer bound to three Fas molecules was generated using knowledge-based protein modeling methods. Point mutations of amino acid residues predicted to affect the receptor-ligand interaction were introduced at three sites. The F275L mutant, mimicking the loss of function murine gld mutation, exhibited a high propensity for aggregation and was unable to bind to Fas. Mutants P206R, P206D, and P206F displayed reduced cytotoxicity toward Fas-positive cells with a concomitant decrease in the binding affinity for the recombinant Fas-immunoglobulin Fc fusion proteins. Although the cytotoxic activity of mutant Y218D was unaltered, mutant Y218R was inactive, correlating with the prediction that Tyr-218 of FasL interacts with a cluster of three basic amino acid side chains of Fas. Interestingly, mutant Y218F could induce apoptosis in murine, but not human cells. |
