Photolabelling of the prostaglandin E 2 receptor in cardiac sarcolemmal vesicles

A [ 3H]azidophenacyl ester of PGE 2 ([ 3H]azido-PGE 2) was synthesized and used to photoaffinity label the protein component of the high affinity PGE 2 binding site in cardiac sarcolemma membrane. Photolysis of the isolated cardiac sarcolemmal vesicles in the presence of [ 3H]azido-PGE 2 resulted in...

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Bibliographic Details
Published in:FEBS letters 1990-06, Vol.265 (1), p.117-120
Main Authors: Michalak, Marek, Wandler, Elayne L., Strynadka, Ken, Lopaschuk, Gary L., Njue, Wilson M., Liu, Hsing-Jang, Olley, Peter M.
Format: Article
Language:English
Subjects:
Cardiac sarcolemma
Photolabelling
Prostaglandin receptor
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Summary:A [ 3H]azidophenacyl ester of PGE 2 ([ 3H]azido-PGE 2) was synthesized and used to photoaffinity label the protein component of the high affinity PGE 2 binding site in cardiac sarcolemma membrane. Photolysis of the isolated cardiac sarcolemmal vesicles in the presence of [ 3H]azido-PGE 2 resulted in the covalent labelling of a protein component that migrated on sodium dodecyl sulfate-polyacrylamide gels with an apparent molecular weight of 100 000. Incorporation of the [ 3H]azido-PGE 2 did not occur in the absence of photolysis. The photolabelling of the 100-kDa protein by [ 3H]azido-PGE 2 was inhibited by excess unlabelled PGE 2 and arido-PGE 2. Specific binding of [ 3H]azido-PGE 2 was displaced by excess unlabelled PGE 2 or azido-PGE 2, but not PGF 2α, 6-keto-PGF 1α or PGD 2. These results indicate that the 100-kDa photoaffinity labelled [ 3H]azido-PGE 2 binding protein contains the binding site for PGE 2 in isolated cardiac sarcolemma membranes.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(90)80898-S