Specific binding and degradation of somatostatin by membrane vesicles from pig gut

M. Weber, T. Cole, J. M. Conlon

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Abstract

Plasma membrane vesicles were prepared from the basolateral face of pig small intestinal epithelial cells and were enriched in the activity of Na + -K + -ATPase (9-fold relative to the cell homogenate) and ranged in size from 0.15 to 0.40 μm diam. Incubation of somatostatin-14 and [ 125 I-Tyr 11 ]-somatostatin-14 with the vesicles at 37°C resulted in rapid proteolytic degradation of the peptides. Metabolites were isolated by reverse-phase high-performance liquid chromatography and identified by amino acid composition. Cleavages between Ala 1 -Gly 2 , Phe 6 -Phe 7 -Trp 8 , and Thr 10 -Phe 11 were observed, indicative of aminopeptidase and endopeptidase action. Degradation was inhibited by 1,10-phenanthroline and by bacitracin, and in the presence of these inhibitors and at 21°C binding of [ 125 I-Tyr 11 ]somatostatin-14 to the vesicles was observed. Binding was inhibited in a concentration-dependent manner by somatostatin-14 (half-maximal inhibition at 2.0 ± 0.1 nM) and by somatostatin-28 (0.8 ± 0.1 nM) but not by structurally unrelated peptides. The rate of degradation of [ 125 I-Leu 8 , D-Trp 22 , Tyr 25 ]somatostatin-28 by basolateral membrane was < 20 fold that of [ 125 I-Tyr 11 ]somatostatin-14 and a two- to three-fold enhanced binding to the vesicles was observed. Analysis of the inhibition of binding of this analogue by somatostatin-28 indicates the presence of single class of binding site with K(d) = 1.3 ± 0.3 nM. Rapid degradation but no specific binding of somatostatin-14 by brush-border membranes was observed. Binding of somatostatins by epithelial cell membranes may be related to the regulation of nutrient absorption by the gut, but the rapid rate of degradation suggests that binding may also serve to limit the release of the peptides into the circulation.

Original languageEnglish
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume250
Issue number5
Publication statusPublished - 1 Jan 1986
Externally publishedYes

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