Photoaffinity labelling of Ca2+ channels with [3H]azidopine
- 9 April 1984
- journal article
- Published by Wiley in FEBS Letters
- Vol. 169 (1) , 112-118
- https://doi.org/10.1016/0014-5793(84)80299-9
Abstract
A 1,4-dihydroypyridine arylazide photoaffinity ligand, [3H]azidopine (50.6 Ci/mmol), has been synthesized. [3H]Azidopine binds reversibly with a Kd of 350 pM to guinea-pig skeletal muscle membranes in the absence of ultraviolet light. The reversible [3H]azidopine binding is inhibited stereoselectively by 1,4-dihydropyridines, phenylalkylamine Ca2+ channel blockers and La3+. Covalent incorporation into membrane proteins after photolysis was investigated by sodium dodecyl sulfate polyacrylamide slab gel electrophoresis. [3H]Azidopine is photoincorporated specifically into a protein of Mr∼145 000. The covalent labelling of the Mr ∼145 ooo band is inhibited stereoselectively by drugs and cations which block the reversible [3H]azidopine binding. It is suggested that [3H]azidopine is photoincorporated into a subunit of the putative Ca2+ channelKeywords
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