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Original Articles

O-Linked Oligosaccharides on Insulin Receptor

  1. Elaine Collier and
  2. Phillip Gorden
  1. Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health Bethesda, Maryland
  1. Address correspondence and reprint requests to Elaine Collier, MD, Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases, Building 10, Room 8S-243, National Institutes of Health, Bethesda, MD 20892.
Diabetes 1991 Feb; 40(2): 197-203. https://doi.org/10.2337/diab.40.2.197
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Abstract

The insulin receptor, an integral membrane glycoprotein, is synthesized as a single-chain precursor that is cleaved to produce two mature subunits, both of which contain N-linked oligosaccharide chains and covalently linked fatty acids. We report that the β-subunit also contains O-linked oligosaccharides. The proreceptor, α-subunit, and β-subunit were labeled with [3H]mannose and [3H]galactose in the presence or absence of an inhibitor of O-linked glycosylation. Tryptic peptides from each component were separated by reverse-phase high-performance liquid chromatography. N- and O-linked oligosaccharide chains were identified on these peptides by specific enzymatic digestions. The proreceptor and α-subunit contained only N-linked oligosaccharides, whereas the β-subunit contained both N- and O-linked oligosaccharides. The O-linked oligosaccharide chains were attached to a single tryptic fraction of the β-subunit, which also contained N-linked chains. This fraction was further localized to the NH2-terminal tryptic peptide of the β-subunit by specific immunoprecipitation with an anti-peptide antibody with specificity for this region. Binding of insulin and autophosphorylation of the β-subunit were not dependent on O-linked glycosylation, because cells grown in the presence of the inhibitor exhibited a normal dose response to insulin. Therefore, the insulin receptor contains O-linked oligosaccharides on the NH2-terminal tryptic peptide of the β-subunit, and these O-linked oligosaccharides are not necessary to the binding or autophosphorylation function of the receptor.

  • Received April 2, 1990.
  • Revision received September 7, 1990.
  • Accepted September 7, 1990.
  • Copyright © 1991 by the American Diabetes Association

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February 1991, 40(2)
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O-Linked Oligosaccharides on Insulin Receptor
Elaine Collier, Phillip Gorden
Diabetes Feb 1991, 40 (2) 197-203; DOI: 10.2337/diab.40.2.197

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O-Linked Oligosaccharides on Insulin Receptor
Elaine Collier, Phillip Gorden
Diabetes Feb 1991, 40 (2) 197-203; DOI: 10.2337/diab.40.2.197
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