Structure and function in rhodopsin: covalent crosslinking of the rhodopsin (metarhodopsin II) -transducin complex- the rhodopsin cytoplasmic face links to the transducin a subunit

Resek, J.F.; Farrens, D.; Khorana, H.Gobind

Proceedings of the National Academy of Sciences of the United States of America 91: 43-7

1994


ISSN/ISBN: 0027-8424
Accession: 009986534

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Abstract
The sites of interaction between rhodopsin and transducin were mapped. Mutant rhodopsin molecules were prepared containing a single reactive cysteine residue per rhodopsin molecule at position 65, 140, 240, or 316 on the cytoplasmic face. The cysteine sulfhydryl group of each of the rhodopsin molecules was linked to a carbene-generating photoactivatable group via a disulfide bond, and this derivative was light-activated at l > 495 nm. Metarhodopsin II was thus formed and found to bind transducin. When the rhodopsin mutant with cysteine at the 240 position was subsequently photoactivated at 355 nm, it was found to covalently crosslink with transducin via the a subunit of transducin.