Carbohydrate determinants involved in both the binding and action of insulin in rat adipocytes.
The insulin receptor apparent affinity was markedly decreased in fat cells treated with lectins specific either for D-galactose (Ricinus communis agglutinin I, RCAI), D-mannose (concanavalin A, Con A, Lens culinaris agglutinin, LCA) or N-acetyl-D-glucosamine (wheat germ agglutinin, WGA), as indicated by a rightward shift of the binding competition curves and almost lineared Scatchard plots. Limulus polyphemus agglutinin (LPA), specific for sialic acid, was ineffective. All lectins enhanced 2-deoxy-D-glucose uptake with relative bioactivities (maximal lectin effect/maximal insulin effect) of 68-86%. Insulin and lectin stimulatory effects were antagonized by specific carbohydrates used as competitors and inhibited by cytochalasin B (70 microM). Maximal effects of insulin and lectins were not additive and were completely abolished in neuraminidase-treated fat cells. Lectins did not affect insulin degradation. These data show that sialylated glycosidic moieties containing D-galactose, D-mannose and N-acetyl-D-glucosamine units are involved in both processes of insulin 'high affinity' binding and activation of glucose transport but are not implicated in hormone degradation. They suggest that N-linked carbohydrate chains of the complex type may be essential for functional insulin receptor and post-receptor systems.
D000117:Acetylglucosamine / Q000502:physiology; D000273:Adipose Tissue / Q000378:metabolism*; D000818:Animals; D002241:Carbohydrates / Q000502:physiology*; D003571:Cytochalasin B / Q000494:pharmacology; D003847:Deoxyglucose / Q000378:metabolism; D005690:Galactose / Q000502:physiology; D007328:Insulin / Q000378:metabolism / Q000494:pharmacology*; D037102:Lectins / Q000494:pharmacology; D008358:Mannose / Q000502:physiology; D009439:Neuraminidase / Q000494:pharmacology; D051381:Rats; D011972:Receptor, Insulin / Q000187:drug effects / Q000378:metabolism*; D013329:Structure-Activity Relationship
D002241:Carbohydrates; D007328:Insulin; D037102:Lectins; D003571:Cytochalasin B; D003847:Deoxyglucose; D011972:Receptor, Insulin; D009439:Neuraminidase; D008358:Mannose; D000117:Acetylglucosamine; D005690:Galactose