Purification of A-subtype pancreatic cholecystokinin receptor by immunoaffinity chromatography.

Abstract:

:So far, no efficient affinity chromatography for CCK receptor purification has been reported that prevented obtention of sequenceable amounts of purified receptor. In this work, 10% of plasma membrane receptor sites were specifically cross-linked with the photoreactive cleavable agonist 125I-ASD-[Thr28, Ahx31]-CCK-25-33, solubilized by NP-40, chromatographied on immobilized wheat germ agglutinin and further immunopurified using anti-CCK antibodies to an overall rate of 3000-3600-fold. Analysis of eluted material demonstrated a protein migrating at Mr 85,000-100,000 and the absence of 35S-labeled impurity. This single and efficient affinity chromatography should provide enough homogeneous receptor protein for microsequence determination and leads to consider immunoaffinity chromatography on immobilized anti-ligand antibodies as a potential tool for purification of membrane receptors.

journal_name

Biochimie

journal_title

Biochimie

authors

Dufresne M,Poirot S,Jimenez J,Vaysse N,Fourmy D

doi

10.1016/0300-9084(92)90039-h

keywords:

subject

Has Abstract

pub_date

1992-02-01 00:00:00

pages

149-51

issue

2

eissn

0300-9084

issn

1638-6183

pii

0300-9084(92)90039-H

journal_volume

74

pub_type

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