Self-association of tissue factor as revealed by chemical crosslinking.
The possible self-association of tissue factor molecules was investigated by treating cells expressing tissue factor with bifunctional cross-linking agents. The two reagents chosen were 3,3'-dithiobis(sulfosuccinimidylpropionate) and sulfosuccinimidyl 2-(p-azidosalicylamido)ethyl-1,3'-dithiopropionate, both of which are membrane-impermeable and thiol-cleavable. A human bladder carcinoma cell line, J82, and a transfected human kidney cell line expressing high amounts of recombinant tissue factor were used in these studies. Exposure of the intact cells to the crosslinking reagents was found to result in the formation of multimeric tissue factor-containing complexes, the extent of which appeared to be dependent upon the amount of tissue factor expressed by the cell. The self-association of tissue factor was prevented in a variant tissue factor molecule harboring a non-homologous transmembrane domain.
Roy
S
S
Department of Cardiovascular Research, Genentech Inc., South San Francisco, California 94080.
Paborsky
L R
LR
Vehar
G A
GA
D000595:Amino Acid Sequence; D001345:Autoradiography; D015153:Blotting, Western; D002478:Cells, Cultured; D003432:Cross-Linking Reagents; D006801:Humans; D007668:Kidney / Q000166:cytology; D008969:Molecular Sequence Data; D010957:Plasmids; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000737:chemistry; D013925:Thromboplastin / Q000737:chemistry* / Q000235:genetics; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000473:pathology
D003432:Cross-Linking Reagents; D011994:Recombinant Proteins; D013925:Thromboplastin