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
Biochimiejournal_title
Biochimieauthors
Dufresne M,Poirot S,Jimenez J,Vaysse N,Fourmy Ddoi
10.1016/0300-9084(92)90039-hkeywords:
subject
Has Abstractpub_date
1992-02-01 00:00:00pages
149-51issue
2eissn
0300-9084issn
1638-6183pii
0300-9084(92)90039-Hjournal_volume
74pub_type
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