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Identification of functional insulin receptors on membranes from an insulin-producing cell line (RINm5F)

Gazzano, H.
Ballotti, R.
Brandenburg, D.
Fehlmann, M.
Van Obberghen, E.
Published in Biochemical Journal. 1985, vol. 226, no. 3, p. 867-872
Abstract Insulin receptors on RINm5F cell membranes (an insulin-producing rat pancreatic cell line) were studied. To study the insulin receptor alpha-subunit, 125I-labelled photoreactive insulin was covalently bound to the membranes in the absence or presence of unlabelled insulin. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis under reducing conditions showed specific labelling of an Mr 130 000 protein. The receptor beta-subunit was studied by using a cell-free phosphorylation assay. Analysis under reducing conditions showed a phosphoprotein of Mr 95 000 whose level of phosphorylation was selectively increased by insulin, and which was specifically immunoprecipitated by antibodies to the insulin receptor. Further, covalent hormone-receptor complexes purified with anti-insulin antibodies were able to undergo autophosphorylation, indicating the existence of operational receptor subunit arrangements. RINm5F cell insulin receptors (and, by analogy, possibly those of native B-cells) thus display structural and functional integrity comparable with those of conventional insulin target cells.
Keywords AnimalsCell LineCell Membrane/metabolismChemical PrecipitationElectrophoresis, Polyacrylamide GelIslets of Langerhans/ metabolismPhosphorylationRatsReceptor, Insulin/immunology/ metabolism
PMID: 3885941
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GAZZANO, H. et al. Identification of functional insulin receptors on membranes from an insulin-producing cell line (RINm5F). In: Biochemical Journal, 1985, vol. 226, n° 3, p. 867-872. https://archive-ouverte.unige.ch/unige:8753

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Deposited on : 2010-07-12

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