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Photolabelling of the prostaglandin E 2 receptor in cardiac sarcolemmal vesicles
Author(s) -
Michalak Marek,
Wandler Elayne L.,
Strynadka Ken,
Lopaschuk Gary L.,
Njue Wilson M.,
Liu Hsing-Jang,
Olley Peter M.
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)80898-s
Subject(s) - sarcolemma , chemistry , prostaglandin e , photoaffinity labeling , affinity label , covalent bond , biochemistry , prostaglandin , receptor , membrane , microbiology and biotechnology , biology , organic chemistry
A [ 3 H]azidophenacyl ester of PGE 2 ([ 3 H]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 [ 3 H]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 [ 3 H]azido‐PGE 2 did not occur in the absence of photolysis. The photolabelling of the 100‐kDa protein by [ 3 H]azido‐PGE 2 was inhibited by excess unlabelled PGE 2 and arido‐PGE 2 . Specific binding of [ 3 H]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 [ 3 H]azido‐PGE 2 binding protein contains the binding site for PGE 2 in isolated cardiac sarcolemma membranes.