Identification and characterization of cellular receptors for the growth regulator, oncostatin M.
Oncostatin M is a polypeptide growth regulator produced by activated T cells and phorbol ester-treated U937 cells. To identify specific cellular receptors for this factor, we have characterized the binding of 125I-labeled oncostatin M to a variety of normal and malignant mammalian cells. Recombinant oncostatin M was labeled with 125I with full retention of growth inhibitory activity on A375 melanoma cells. 125I-Oncostatin M bound to sensitive cells in a time- and temperature-dependent fashion. Binding was specifically inhibited by unlabeled native or recombinant oncostatin M, but not by other polypeptide growth factors tested. Binding to human leukemic and normal blood cells was generally less than to nonhematopoietic cells. With four different cell lines, maximal growth inhibition by oncostatin M was achieved at less than maximal binding site occupancy. Scatchard graphs of direct binding data were curvilinear and indicated that 125I-oncostatin M bound with higher apparent affinity at lower 125I-oncostatin M concentrations. Using a two binding site model, affinity constants of Kd1 = 11 +/- 11 pM and Kd2 = 1000 +/- 380 pM were extrapolated from binding data with A375 cells, and values of Kd1 = 3 +/- 2 pM and Kd2 = 400 +/- 44 pM from A549 cells. The major 125I-oncostatin M binding species in a number of mammalian cell lines was identified by chemical cross-linking as a specific protein(s) of Mr = 150,000-160,000. 125I-Oncostatin M was internalized (t1/2 = 30 min) and degraded subsequent to binding to a responsive cell line.(ABSTRACT TRUNCATED AT 250 WORDS)
Linsley
P S
PS
Oncogen, Seattle, Washington 98121.
Bolton-Hanson
M
M
Horn
D
D
Malik
N
N
Kallestad
J C
JC
Ochs
V
V
Zarling
J M
JM
Shoyab
M
M
D000818:Animals; D000970:Antineoplastic Agents; D001667:Binding, Competitive; D002448:Cell Adhesion; D002455:Cell Division / Q000187:drug effects; D003110:Colonic Neoplasms / Q000378:metabolism; D003432:Cross-Linking Reagents; D004285:Dogs; D000882:Haplorhini; D007457:Iodine Radioisotopes; D008175:Lung Neoplasms / Q000378:metabolism; D008545:Melanoma / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D053683:Oncostatin M; D010455:Peptides / Q000378:metabolism* / Q000494:pharmacology; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013388:Succinimides; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D003432:Cross-Linking Reagents; D007457:Iodine Radioisotopes; C508351:OSM protein, human; C508352:Osm protein, mouse; D010455:Peptides; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D013388:Succinimides; D053683:Oncostatin M; C019358:disuccinimidyl suberate