CHARACTERIZATION OF A VASOACTIVE INTESTINAL PEPTIDE-SENSITIVE ADENYLATE-CYCLASE IN RAT INTESTINAL EPITHELIAL-CELL MEMBRANES

被引:78
作者
AMIRANOFF, B
LABURTHE, M
DUPONT, C
ROSSELIN, G
机构
[1] Unite de Recherche de Diabetologie et d'Etudes Radio-Immunologiques des Hormones Proteiques (INSERM,, Hôpital Saint-Antoine, 75571 Paris Cedex 12, 184, rue du Faubourg Saint-Antoine
关键词
D O I
10.1016/0304-4165(78)90321-5
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A vasoactive intestinal peptide-sensitive adenylate cyclase in intestinal epithelial cell membranes was characterized. Stimulation of adenylate cyclase activity was a function of vasoactive intestinal peptide concentration over a range of 1 · 10-10-1 · 10-7 M and was increased six-times by a maximally stimulating concentration of vasoactive intestinal peptide. Half-maximal stimulation was observed with 4.1 ± 0.7 nM vasoactive intestinal peptide. Fluoride ion stimulated adenylate cyclase activity to a higher extent than did vasoactive intestinal peptide. Under standard assay conditions, basal, vasoactive inteetinal peptide- and fluoride-stimulated adenylate cyclase activities were proportional to time of incubation up to 15 min and to membrane concentration up to 60 μg protein per assay. The vasoactive intestinal peptide-sensitive enzyme required 5-10 mM Mg2+ and was inhibited by 1 · 10-5 M Ca2+. At sufficiently high concentrations, both ATP (3 mM) and Mg2+ (40 mM) inhibited the enzyme. Secretin also stimulated the adenylate cyclase activity from intestinal epithelial cell membranes but its effectiveness was 1/1000 that of vasoactive intestinal peptide. Prostaglandins E1 and E2 at 1 · 10-5 M induced a two-fold increase of cyclic AMP production. Vasoactive intestinal peptide was the most potent stimulator of adenylate cyclase activity, suggesting an important physiological role of this peptide in the cyclic AMP-dependent regulation of the intestinal epithelial cell function. © 1978.
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页码:474 / 481
页数:8
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