Complete glycosylation of the insulin and insulin-like growth factor I receptors is not necessary for their biosynthesis and function. Use of swainsonine as an inhibitor in IM-9 cells

J Biol Chem. 1986 Jan 15;261(2):970-5.

Abstract

Swainsonine, an indolizidine alkaloid which is a potent inhibitor of the Golgi enzyme, mannosidase II, leads to the production of incompletely processed glycoproteins lacking complex type oligosaccharides. This inhibitor has been used to examine the importance of terminal sugar groups in the biosynthesis and function of both the insulin receptor and the insulin-like growth factor I receptor. IM-9 cells were metabolically labeled using [35S]methionine and the two receptors were independently immunoprecipitated using specific monoclonal antibodies. The incompletely processed receptors have slightly lower molecular weights and contain hybrid rather than complex type oligosaccharides as indicated by their sensitivity to endoglycosidase H and neuraminidase. Both receptors made in the presence of swainsonine are still autophosphorylated in the presence of the respective hormone. The insulin receptor made in the presence of the inhibitor can be affinity labeled at the cell surface using 125I-insulin and disuccinimidyl suberate cross-linking; there is also no significant difference in its affinity for insulin. These results suggest that for the insulin and insulin-like growth factor I receptors to be synthesized, processed, and function normally, they do not require all of the sugars which are normally added in the terminal stages of glycosylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Affinity Labels / metabolism
  • Alkaloids / pharmacology*
  • Animals
  • Carbohydrate Metabolism*
  • Cell Line
  • Golgi Apparatus / enzymology
  • Mannosidases / antagonists & inhibitors
  • Methionine / metabolism
  • Molecular Weight
  • Neuraminidase / metabolism
  • Phosphorylation
  • Receptor, Insulin / biosynthesis
  • Receptor, Insulin / metabolism*
  • Receptors, Cell Surface / biosynthesis
  • Receptors, Cell Surface / metabolism*
  • Receptors, Somatomedin
  • Swainsonine

Substances

  • Affinity Labels
  • Alkaloids
  • Receptors, Cell Surface
  • Receptors, Somatomedin
  • Methionine
  • Receptor, Insulin
  • Mannosidases
  • mannosyl-oligosaccharide 1,3 - 1,6-alpha-mannosidase
  • Neuraminidase
  • Swainsonine