Interaction of ovine somatomedin-C/IGF-I and IGF-I with specific IGF-I receptors on cultured muscle-derived fibroblasts.
Binding of 125I-insulin-like growth factor-I and 125I-ovine somatomedin-C/IGF-I to monolayer cultures of muscle-derived ovine fibroblasts is described. Preliminary competitive binding experiments indicate that ovine fibroblasts possess independent cell surface receptors for IGF-I. Affinity of rIGF-II for IGF-I binding sites is minimal; rIGF-II binds to Type I IGF receptors at 1/1000 the strength of IGF-I. Insulin binds to the Type I IGF receptor at 1/100 the strength of IGF-I, whereas ovine somatomedin-C/IGF-I displays equivalent IGF-I binding as evidenced by overlapping competition of ovine somatomedin-C/IGF-I for 125I-IGF-I binding sites. Results from disuccinimidyl suberate cross-linking of 125I-IGF-I to muscle-derived ovine fibroblasts in the presence of related polypeptides verified the competitive binding data. Under reducing conditions, 125I-IGF-I: receptor complexes migrated to a relative molecular weight of approximately 135,000 daltons. Specific 125I-IGF-I binding was completely inhibited by 10(-8) mol/l IGF-I, 7.2 x 10(-8) mol/l ovine somatomedin-C/IGF-I, and 10(-6) mol/l insulin and partially inhibited by 7.2 x 10(-9) mol/l ovine somatomedin-C/IGF-I and 6.5 x 10(-8) mol/l rIGF-II. 125I-ovine somatomedin-C/IGF-I: receptor complexes also migrated at a relative molecular weight of 135,000 daltons. No migratory band was observed at 250,000 to 260,000 daltons with either 125I-IGF-I or 125I-ovine somatomedin-C/IGF-I indicating that little labelled moiety bound to the Type II IGF receptor.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D001345:Autoradiography; D001667:Binding, Competitive; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism*; D007334:Insulin-Like Growth Factor I / Q000378:metabolism*; D008297:Male; D011972:Receptor, Insulin / Q000378:metabolism*; D017451:Receptors, Somatomedin; D012756:Sheep; D013002:Somatomedins / Q000378:metabolism*
D017451:Receptors, Somatomedin; D013002:Somatomedins; D007334:Insulin-Like Growth Factor I; D011972:Receptor, Insulin