Expression of atypical and classical insulin receptors in Chinese hamster ovary cells transfected with cloned cDNA for the human insulin receptor

• Published in: Endocrinology (Philadelphia) Vol. 127; no. 3; p. 1301
• Main Authors: Jonas, H A, Eckardt, G S, Clark, S
• Format: Journal Article
• Language: English
• Published: United States 01.09.1990
• Subjects: Animals; Binding, Competitive; Cell Line; Cloning, Molecular; Cricetinae; DNA - genetics; Gene Expression; Humans; Insulin - metabolism; Insulin - pharmacology; Insulin-Like Growth Factor I - metabolism; Insulin-Like Growth Factor II - metabolism; Protein Processing, Post-Translational; Receptor, Insulin - genetics; Receptor, Insulin - metabolism; Somatomedins - metabolism; Transfection
• DOI: 10.1210/endo-127-3-1301
• ISSN: 0013-7227

Human placenta and IM-9 lymphocytes contain subpopulations of atypical insulin receptors which differ from classical insulin receptors in their higher binding affinity for insulin-like growth factors I and II (IGF-I and IGF-II). Both types of insulin receptors may be derived from different but related genes, or may represent alternative post-translational modifications of the same gene product. To test these possibilities, we have examined the IGF binding characteristics of the human insulin receptors expressed in Chinese hamster ovary (CHO) cells which had been stably transfected with cloned human insulin receptor cDNA (CHO-T cells). The parent CHO cells contained 3 x 10(3) rodent insulin receptors/cell, and the CHO-T cells, 2.0 x 10(6) human insulin receptors/cell. Competition binding studies showed that the binding of [125I]IGF-I and [125I]multiplication stimulating activity (MSA/rat IGF-II) to parent CHO cells was primarily to type I and II IGF receptors, which cross-react poorly or not at all with insulin. However, competition binding studies with CHO-T cells showed that [125I]IGF-I binding was displaced 60-70%, and [125I]MSA binding, 50-55%, by low concentrations of insulin (20 ng/ml) and no further by higher concentrations of insulin (500 ng/ml). The insulin-insensitive IGF binding sites corresponded to the rodent type I and II IGF receptors; the insulin-sensitive IGF binding sensitive sites resembled the human atypical insulin receptors in that they bound IGF-I and MSA with moderately high affinity and reacted with insulin, MSA, and IGF-I in that order of potency. Atypical insulin receptors were also demonstrated by insulin-sensitive [125I]IGF-I and [125I]MSA binding to solubilized CHO-T proteins adsorbed to microtiter wells coated with monoclonal antibodies specific for the human insulin receptor. These results suggest that atypical human insulin receptors are generated by differential post-translational processing of the same gene product as classical human insulin receptors.