Reference : Purification and molecular cloning of the APO-1 cell surface antigen, a member of the...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
Purification and molecular cloning of the APO-1 cell surface antigen, a member of the tumor necrosis factor/nerve growth factor receptor superfamily. Sequence identity with the Fas antigen
Oehm, A. [> >]
Behrmann, Iris mailto [University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit >]
Falk, W. [> >]
Pawlita, M. [> >]
Maier, G. [> >]
Klas, C. [> >]
Li-Weber, M. [> >]
Richards, S. [> >]
Dhein, J. [> >]
Trauth, B. C. [> >]
Journal of Biological Chemistry
American Society for Biochemistry and Molecular Biology
Yes (verified by ORBilu)
[en] Amino Acid Sequence ; Humans ; Membrane Proteins ; Molecular Sequence Data ; Multigene Family ; Nerve Growth Factors ; Receptors, Cell Surface ; Receptors, Nerve Growth Factor ; Receptors, Tumor Necrosis Factor ; Sequence Homology, Nucleic Acid ; Enzyme-Linked Immunosorbent Assay ; Electrophoresis, Polyacrylamide Gel ; Antigens, CD95 ; Antigens, Neoplasm ; Antigens, Surface ; Base Sequence ; Blotting, Western ; Cell Death ; Cloning, Molecular ; Cross-Linking Reagents ; DNA ; Tumor Necrosis Factor-alpha
[en] The APO-1 antigen as defined by the mouse monoclonal antibody anti-APO-1 was previously found to be expressed on the cell surface of activated human T and B lymphocytes and a variety of malignant human lymphoid cell lines. Cross-linking of the APO-1 antigen by anti-APO-1 induced programmed cell death, apoptosis, of APO-1 positive cells. To characterize the APO-1 cell surface molecule and to better understand its role in induction of apoptosis, the APO-1 protein was purified to homogeneity from membranes of SKW6.4 B lymphoblastoid cells by solubilization with sodium deoxycholate, affinity chromatography with anti-APO-1 antibody, and reversed phase high performance liquid chromatography. Each purification step was followed by an APO-1-specific solid phase enzyme-linked immunosorbent assay using the monoclonal antibody anti-APO-1. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the APO-1 antigen was found to be a membrane glycoprotein of 48-kDa. Endoproteinase-cleaved peptides of the APO-1 protein were subjected to amino acid sequencing, and corresponding oligonucleotides were used to identify a full-length APO-1 cDNA clone from an SKW6.4 cDNA library. The deduced amino acid sequence of APO-1 showed sequence identity with the Fas antigen, a cysteine-rich transmembrane protein of 335 amino acids with significant similarity to the members of the tumor necrosis factor/nerve growth factor receptor superfamily. The APO-1 antigen was expressed upon transfection of APO-1 cDNA into BL60-P7 Burkitt's lymphoma cells and conferred sensitivity towards anti-APO-1-induced apoptosis to the transfectants.

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