Identification and characterization of insulin receptors in basolateral membranes of dog intestinal mucosa.
Little is known about hormonal regulation of substrate transport and metabolism in the mucosal lining of the small intestine. Because insulin regulates these functions in other tissues by binding to its receptor, we have investigated the presence of insulin receptors in canine small intestinal mucosa with basolateral membranes (BLM) and brush border membranes (BBM) prepared by sorbitol density centrifugation. A14-[125I]iodoinsulin was used to study binding and structural characteristics of specific insulin receptors in BLM. Analysis of receptors in BLM identified binding sites with high affinity (Kd 88 pM) and low capacity (0.4 pmol/mg protein) as well as with low affinity (Kd 36 nM) and high capacity (4.7 pmol/mg protein). Binding was time, temperature, and pH dependent, and 125I-labeled insulin dissociation was enhanced in the presence of unlabeled insulin. Cross-reactivity of these receptors to proinsulin, IGF-II, and IGF-I was 4, 1.8, and less than 1%, respectively. Covalent cross-linking of labeled insulin to BLM insulin receptors with disuccinimidyl suberate revealed a single 135,000-Mr band that was completely inhibited by unlabeled insulin. There was a 16-fold greater specific binding of insulin to BLM (39.0 +/- 2.4%) than to BBM (2.5 +/- 0.6%). These results demonstrate the presence of a highly specific receptor for insulin on the vascular, but not the luminal, surface of the small intestinal mucosa in dogs, and suggest that insulin may play an important role in the regulation of gastrointestinal physiology.
Gingerich
R L
RL
Edward Mallinckrodt Department of Pediatrics, Washington University School of Medicine, St Louis, Missouri 63178.
Gilbert
W R
WR
Comens
P G
PG
Gavin
J R
JR
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D004285:Dogs; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000031:analogs & derivatives / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism*; D007421:Intestine, Small / Q000378:metabolism; D007457:Iodine Radioisotopes; D007700:Kinetics; D008871:Microvilli / Q000378:metabolism; D011972:Receptor, Insulin / Q000378:metabolism*; D013696:Temperature
D007328:Insulin; D007457:Iodine Radioisotopes; C008147:insulin, iodo-; D011972:Receptor, Insulin