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Scientific article
English

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

Contributors

Gazzano, H.; Halban, Philippe A.; Prentki, Marc; Ballotti, R.; Brandenburg, D.; Fehlmann, M.; Van Obberghen, E.

Published in

Biochemical journal, vol. 226, no. 3, p. 867-872

Publication date

1985

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

  • Animals
  • Cell Line
  • Cell Membrane/metabolism
  • Chemical Precipitation
  • Electrophoresis, Polyacrylamide Gel
  • Islets of Langerhans/ metabolism
  • Phosphorylation
  • Rats
  • Receptor, Insulin/immunology/ metabolism

UNIGE affiliation entities

Citation (ISO format)

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. doi: 10.1042/bj2260867

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Additional URL for this publication

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1144788/pdf/biochemj00307-0234.pdf

Journal ISSN

0264-6021
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