Intracellular (Golgi) receptors for insulinlike peptides in rat liver

Abstract

An insulin-like peptide (ILA) was isolated from human serum. The binding of 125I-labeled ILA was studied in subcellular fractions from rat liver and found to be much greater in microsomes than purified plasmalemma. The high level of microsomal binding was due to a particular enrichment of binding sites in Golgi elements. Binding to Golgi was time and temperature-dependent and was augmented by an increase of either subcellular fraction or 125I-labeled ILA in accordance with a mass action process. Degradation of 125I-labeled ILA was greatest in the Golgi vesicle fraction and was reduced by incubation at 4.degree. C. Bound 125I-labeled ILA could be eluted and was found to retain integrity. Binding was pH dependent with a broad optimum at pH 7.7-8.5. Dissociation of bound 125I-labeled ILA was time and temperature-dependent. It was greater at 37.degree. C than 4.degree. C, and was uninfluenced by unlabeled ILA. The ILA receptor was stable at 4.degree. C but was markedly decreased by preincubation at 37.degree. C. The binding of 125I-labeled ILA was inhibited by unlabeled ILA and related insulinlike peptides (the insulinlike growth factors, IGF-1 and IGF-2) in a dose-dependent manner. Insulin and its analogs had only a partially inhibitory effect and structurally unrelated peptides were without inhibitory efficacy. In contrast, ILA and IGF-1 and IGF-2 inhibited 125I-labeled insulin binding to its receptors in a dose-dependent fashion. These observations identify a receptor for insulinlike peptides in the Golgi elements of rat liver. It is distinct from the insulin receptor previously observed in these elements. The dual interaction of ILA and other insulinlike peptides with both the insulin receptor and their own unique receptor constitutes the presumed biochemical basis for the 2 types of action effected by this family of peptides, namely, an effect on metabolism comparable to insulin and an effect on cellular anabolism and growth.