Substitution of arginine for histidine at position 209 in the alpha-subunit of the human insulin receptor. A mutation that impairs receptor dimerization and transport of receptors to the cell surface

J Biol Chem. 1991 Nov 5;266(31):21224-31.

Abstract

Receptors for insulin and epidermal growth factor contain cysteine-rich domains in the extracellular portion of the molecule. His209 (insulin receptor numbering system) is 1 of 2 amino acid residues that are identically conserved in the cysteine-rich domains of insulin receptors, epidermal growth factor receptors, and other homologous receptors. Previously, we have identified a patient with leprechaunism who is homozygous for a mutation substituting Arg for His209 in the insulin receptor gene (Kadowaki, T., Kadowaki, H., Rechler, M. M., Serrano-Rios, M., Roth, J., Gorden, P., and Taylor, S. I. (1990) J. Clin. Invest. 86, 254-264). In this investigation, the Arg209 mutant receptor was expressed by transfection of mutant cDNA into NIH-3T3 cells. The mutation impairs several steps in the post-translational processing of the insulin receptor:dimerization of 190-kDa proreceptors into a disulfide linked species, proteolytic cleavage of the proreceptor into alpha- and beta-subunits, and terminal processing of the high mannose form of N-linked oligosaccharide into complex carbohydrate. In addition, the defects in post-translational processing within the endoplasmic reticulum and Golgi apparatus are associated with a marked inhibition in transport of receptors to the plasma membrane. Nevertheless, a small number (approximately 10%) of the receptors are transported to the cell surface. These receptors on the cell surface bind insulin with normal affinity and have normal tyrosine kinase activity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells
  • Amino Acid Sequence
  • Animals
  • Arginine / physiology
  • Base Sequence
  • Biological Transport
  • Cell Membrane / metabolism
  • DNA Mutational Analysis
  • Histidine / physiology
  • Insulin / metabolism
  • Insulin Resistance / genetics*
  • Macromolecular Substances
  • Mice
  • Molecular Sequence Data
  • Phosphorylation
  • Protein Conformation
  • Protein Processing, Post-Translational
  • Protein-Tyrosine Kinases / metabolism
  • Receptor, Insulin / genetics
  • Receptor, Insulin / metabolism*
  • Structure-Activity Relationship
  • Transfection

Substances

  • Insulin
  • Macromolecular Substances
  • Histidine
  • Arginine
  • Protein-Tyrosine Kinases
  • Receptor, Insulin