In:
European Journal of Biochemistry, Wiley, Vol. 191, No. 1 ( 1990-07), p. 141-146
Abstract:
The present study was undertaken to characterize the immune recognition of pancreatic cholecystokinin receptor by an anti‐cholecystokinin antibody. Cholecystokinin receptor from pancreatic plasma membranes was photoaffinity labelled using the specific, cleavable probe 125 I‐labelled 2‐( p ‐azidosalicylamido)‐1,3‐dithiopro‐pionate‐[Thr 28 , Ahx 31 ]CCK(25–33) [CCK(25–33) is the C‐terminal nonapeptide of the 33‐amino‐acid form of cholecystokinin] . Labelled receptor was then solubilized and subsequently prepurified on immobilized wheat‐germ agglutinin. The C‐terminal‐directed anti‐cholecystokinin serum (8E) specifically immunoprecipitated a fraction of affinity‐labelled cholecystokinin receptor which was identified at M r 85000–100000 on SDS/PAGE. The binding affinity of antiserum 8E for covalently labelled cholecystokinin receptor was lower (K d 0.11 ± 0.02 nM) than for cholecystokinin (K d 3.65 ± 0.55 pM). The compound L364‐718, an A‐subtype cholecystokinin‐receptor antagonist did not interfere with the immune recognition of cholecystokinin. However, the recognition of affinity‐labelled cholecystokinin receptor was enhanced as a result of an increasing availability of cholecystokinin molecules. Indeed, the amount of immunoprecipitated receptor was doubled in the presence of 10 μM L364–718. This study offers the possibility of using an anti‐cholecystokinin antibody for cholecystokinin‐receptor purification and demonstrates that prepurified affinity‐labelled cholecystokinin receptor retains A‐subtype specificity.
Type of Medium:
Online Resource
ISSN:
0014-2956
,
1432-1033
DOI:
10.1111/ejb.1990.191.issue-1
DOI:
10.1111/j.1432-1033.1990.tb19103.x
Language:
English
Publisher:
Wiley
Publication Date:
1990
detail.hit.zdb_id:
1398347-7
detail.hit.zdb_id:
1464377-7
SSG:
12
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