Biochemical analysis of the epithelin receptor.
Epithelin 1 and 2 are cysteine-rich proteins that act as growth modulators of epithelial cells. In this report, we have characterized the epithelins receptors of MDA-MB-468 human breast carcinoma cells using both binding and cross-linking techniques. Equilibrium binding studies of iodinated epithelin 1 indicated that two classes of binding sites are expressed at the surface of breast carcinoma cells. The high affinity sites had a dissociation constant of approximately 2 x 10(-10) M, with approximately 290 receptors/cell. The low affinity sites had a dissociation constant of approximately 10(-8) M, with approximately 32,000 receptors/cell. Binding of iodinated epithelin 1 was specifically inhibited by unlabeled epithelin 1, 2, or 3, but not by other growth factors tested. We also performed competition binding studies of 125I-epithelin 1 to cell surface receptors in the presence of unlabeled epithelin 1, 2, or 3. Binding results analyzed by the method of Scatchard suggested that all three epithelins interact with a same receptor. A 140-145-kDa epithelin 1-binding protein complex was identified by chemical cross-linking of 125I-epithelin 1 to breast cancer cells. Formation of such a complex was prevented by coincubation of 125I-epithelin 1 with an excess of unlabeled epithelin 1, 2, or 3.
Culouscou
J M
JM
Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, Washington 98121.
Carlton
G W
GW
Shoyab
M
M
D001665:Binding Sites; D001667:Binding, Competitive; D001943:Breast Neoplasms; D002455:Cell Division / Q000187:drug effects; D016207:Cytokines / Q000494:pharmacology; D005260:Female; D000077155:Granulins; D006131:Growth Inhibitors / Q000378:metabolism* / Q000494:pharmacology*; D006133:Growth Substances / Q000378:metabolism* / Q000494:pharmacology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007700:Kinetics; D011956:Receptors, Cell Surface / Q000302:isolation & purification / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D016207:Cytokines; D000077155:Granulins; D006131:Growth Inhibitors; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D011956:Receptors, Cell Surface