Photolabelling of the prostaglandin E2 receptor in cardiac sarcolemmal vesicles

FEBS Letters

Lopaschuk, Gary L.¹  Njue, Wilson M.²  Liu, Hsing-Jang²  Strynadka, Ken¹  Wandler, Elayne L.¹  Michalak, Marek¹  Olley, Peter M.¹ 

[1] Cardiovascular Disease Research Group, University of Alberta, Edmonton, Alta., Canada T6G 2S2;Department of Chemistry, University of Alberta, Edmonton, Alta., Canada T6G 2S2

关键词: Prostaglandin receptor;    Cardiac sarcolemma;    Photolabelling;    PG;    prostaglandin;    SL;    sarcolemma;    SDS-PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;    Hepes;    4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid;    azido-PGE2;    azidophenacyl ester of PGE2;   

DOI  :  10.1016/0014-5793(90)80898-S

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A [³H]azidophenacyl ester of PGE₂ ([³H]azido-PGE₂) was synthesized and used to photoaffinity label the protein component of the high affinity PGE₂ binding site in cardiac sarcolemma membrane. Photolysis of the isolated cardiac sarcolemmal vesicles in the presence of [³H]azido-PGE₂ 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-PGE₂ did not occur in the absence of photolysis. The photolabelling of the 100-kDa protein by [³H]azido-PGE₂ was inhibited by excess unlabelled PGE₂ and arido-PGE₂. Specific binding of [³H]azido-PGE₂ was displaced by excess unlabelled PGE₂ or azido-PGE₂, but not PGF_2α, 6-keto-PGF_1α or PGD₂. These results indicate that the 100-kDa photoaffinity labelled [³H]azido-PGE₂ binding protein contains the binding site for PGE₂ in isolated cardiac sarcolemma membranes.

【 授权许可】

Unknown