Reference - Detail
RRC ID | 67354 |
---|---|
Author | Suda T, Nagata S. |
Title | Purification and characterization of the Fas-ligand that induces apoptosis. |
Journal | J Exp Med |
Abstract |
Fas is a 45-kD cell surface protein belonging to the tumor necrosis factor/nerve growth factor receptor family, and transduces the signal for apoptosis. The cytotoxic T lymphocyte (CTL) hybridoma, PC60-d10S requires the presence of Fas on target cells to induce cytolysis in target cells. This CTL cell line was weakly but specifically stained by a chimeric protein that consisted of the extracellular domain of mouse Fas and the Fc portion of human immunoglobulin G1 (mFas-Fc). Moreover, mFas-Fc inhibited the cytotoxic activity of PC60-d10S. Sublines of d10S that were stained intensively by mFas-Fc were isolated by repetitive fluorescence-activated cell sorter sorting. A cell-surface protein of about 40 kD was specifically precipitated by mFas-Fc from the lysates of these sublines. This protein was homogeneously purified by sequential affinity chromatographies using mFas-Fc and concanavalin A beads. The purified protein exhibited cytotoxic activity against cells expressing Fas but not to the cells which do not express Fas. These results indicated that the 40-kD membrane glycoprotein expressed on PC60-d10S cells is the Fas-ligand that induces the apoptotic signal by binding to Fas. |
Volume | 179(3) |
Pages | 873-9 |
Published | 1994-3-1 |
DOI | 10.1084/jem.179.3.873 |
PMID | 7509364 |
PMC | PMC2191418 |
MeSH | Animals Antigens, Surface / biosynthesis Antigens, Surface / metabolism* Apoptosis* Base Sequence Chromatography, Affinity Concanavalin A DNA Primers Electrophoresis, Polyacrylamide Gel Humans Hybridomas / immunology Immunoglobulin Fc Fragments / analysis Immunoglobulin Fc Fragments / metabolism Immunoglobulin G / analysis Immunoglobulin G / metabolism Membrane Glycoproteins / isolation & purification* Membrane Glycoproteins / metabolism Membrane Glycoproteins / physiology Membrane Proteins / isolation & purification* Membrane Proteins / metabolism Mice Molecular Sequence Data Molecular Weight Polymerase Chain Reaction Recombinant Fusion Proteins / analysis Recombinant Fusion Proteins / biosynthesis Recombinant Fusion Proteins / metabolism Signal Transduction T-Lymphocytes, Cytotoxic / immunology* fas Receptor |
IF | 11.743 |
Resource | |
DNA material | pEF-Fc (RDB18974) |