Affinity-labeled plasma somatomedin-C/insulinlike growth factor I binding proteins. Evidence of growth hormone dependence and subunit structure.

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Abstract

By using disuccinimidyl suberate, we have covalently cross-linked 125I-labeled somatomedin-C (Sm-C)/insulinlike growth factor I to specific binding proteins in human plasma. In unfractionated plasma samples from normal and acromegalic donors, 125I-Sm-C binding-protein complexes with relative molecular weights (Mr) of 160,000, 135,000, 110,000, 80,000, 50,000, 43,000-35,000, and 28,000-24,000 were consistently observed. In contrast, the 43,000-35,000-mol wt species were frequently the only specific complexes observed in hypopituitary plasma and were consistently more intensely labeled in such samples. Reduction of samples with beta-mercaptoethanol did not alter the electrophoretic pattern of these 125I-Sm-C binding-protein complexes. All Sm-C binding proteins, with the exception of the 43,000-35,000-mol wt complex, were adsorbed by concanavalin A-Sepharose. When acromegalic or normal plasma was fractionated on a Sephadex G-200 column and affinity labeled, the same complexes that were adsorbed by concanavalin A were found in fractions that eluted near the gamma-globulin peak. On the other hand, the 43,000-35,000-mol wt complex consistently eluted in size-appropriate fractions near the albumin peak. These data suggest that the growth hormone (GH)-dependent Sm-C binding protein, represented by the 160,000-mol wt complex, is in some way composed of smaller species, i.e., the 135,000-, 110,000-, 80,000-, 50,000-, and 28,000-24,000-mol wt complexes. Acid incubation of plasma prior to Sephadex G-200 chromatography results in the elimination of specific 125I-Sm-C binding-protein complexes which elute near gamma-globulin and a concurrent increase in the labeling intensity of the 28,000-24,000-mol wt complexes. We speculate, therefore, that each of the GH-dependent Sm-C binding-protein complexes represents an oligomer composed of 28,000-24,000-mol wt protomers. The 43,000-35,000-mol wt species is not dependent upon GH and appears to represent a different type of Sm-C binding protein.

Authors

J R Wilkins, A J D'Ercole