Oehm, A. and Behrmann, I. and Falk, Werner and Pawlita, M. and Maier, G. and Klas, C. and Li-Weber, M. and Richards, S. and Dhein, J. and Trauth, B. C.
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.
The Journal of biological chemistry 267 (15), pp. 10709-10715.
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.
|Institutions:|| Medicine > Lehrstuhl für Innere Medizin I|
|Amino Acid Sequence||MESH|
|Antigens, Neoplasm/isolation & purification||MESH|
|Electrophoresis, Polyacrylamide Gel||MESH|
|Enzyme-Linked Immunosorbent Assay||MESH|
|Membrane Proteins/isolation & purification||MESH|
|Molecular Sequence Data||MESH|
|Nerve Growth Factors/metabolism||MESH|
|Receptors, Cell Surface/genetics||MESH|
|Receptors, Nerve Growth Factor||MESH|
|Receptors, Tumor Necrosis Factor||MESH|
|Sequence Homology, Nucleic Acid||MESH|
|Tumor Necrosis Factor-alpha/metabolism||MESH|
|Subjects:||600 Technology > 610 Medical sciences Medicine|
|Refereed:||Yes, this version has been refereed|
|Created at the University of Regensburg:||Unknown|
|Deposited On:||08 Sep 2010 05:17|
|Last Modified:||20 Jul 2011 22:36|