Orientation of retinal in bovine rhodopsin determined by cross-linking using a photoactivatable analog of 11-cis-retinal
- PMID: 2144289
Orientation of retinal in bovine rhodopsin determined by cross-linking using a photoactivatable analog of 11-cis-retinal
Abstract
A photoactivatable analog of 11-cis-retinal has been used to probe the orientation of retinal in bovine rhodopsin. The analog binds to the opsin to regenerate a chromophore with lambda max at 458 nm. The linkage site of the analog to the opsin was confirmed to be Lys-296 as in 11-cis-retinal rhodopsin. The analog-reconstituted rhodopsin activated transducin and was phosphorylated by rhodopsin kinase on illumination. On photolysis of rhodopsin containing the radioactively labeled analog at 365 nm at -15 degrees C, 20-25% of the analog was covalently linked to the protein. Proteolysis of the labeled protein and characterization of the appropriate peptides showed that cross-linking of the analog was predominantly to helices C or F. When analog reconstituted rhodopsin in rod outer segments was photolyzed, cross-linking was predominantly to helix C. However, when analog-reconstituted rhodopsin, purified in lauryl maltoside, was photolyzed, labeling occurred mainly in helix F. Sequence analysis showed major sites of cross-linking to be Phe-115, Ala-117, Glu-122, Trp-126, and Ser-127 in helix C while Trp-265 was the major site in helix F. The results suggest that the beta-ionone ring of retinal orients toward helices C and F.
Similar articles
-
Regeneration of bovine and octopus opsins in situ with natural and artificial retinals.Biochemistry. 1989 Mar 21;28(6):2732-9. doi: 10.1021/bi00432a055. Biochemistry. 1989. PMID: 2525050
-
9,13-dicis-rhodopsin and its one-photon-one-double-bond isomerization.Biochemistry. 1988 Aug 23;27(17):6495-9. doi: 10.1021/bi00417a044. Biochemistry. 1988. PMID: 2975508
-
Light-stable rhodopsin. II. An opsin mutant (TRP-265----Phe) and a retinal analog with a nonisomerizable 11-cis configuration form a photostable chromophore.J Biol Chem. 1992 Apr 5;267(10):6770-5. J Biol Chem. 1992. PMID: 1532391
-
Photophysiological functions of visual pigments.Adv Biophys. 1984;17:5-67. doi: 10.1016/0065-227x(84)90024-8. Adv Biophys. 1984. PMID: 6242325 Review.
-
Structural approaches to understanding retinal proteins needed for vision.Curr Opin Cell Biol. 2014 Apr;27:32-43. doi: 10.1016/j.ceb.2013.11.001. Epub 2013 Nov 28. Curr Opin Cell Biol. 2014. PMID: 24680428 Free PMC article. Review.
Cited by
-
Chromophore structural changes in rhodopsin from nanoseconds to microseconds following pigment photolysis.Proc Natl Acad Sci U S A. 1997 Aug 5;94(16):8557-62. doi: 10.1073/pnas.94.16.8557. Proc Natl Acad Sci U S A. 1997. PMID: 9238015 Free PMC article.
-
Automated method for modeling seven-helix transmembrane receptors from experimental data.Biophys J. 1995 Dec;69(6):2419-42. doi: 10.1016/S0006-3495(95)80112-8. Biophys J. 1995. PMID: 8599649 Free PMC article.
-
Differential dynamics in the G protein-coupled receptor rhodopsin revealed by solution NMR.Proc Natl Acad Sci U S A. 2004 Mar 9;101(10):3409-13. doi: 10.1073/pnas.0308713101. Epub 2004 Feb 27. Proc Natl Acad Sci U S A. 2004. PMID: 14990789 Free PMC article.
-
Identification of transmembrane tryptic peptides of rhodopsin using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.Protein Sci. 1997 Apr;6(4):816-24. doi: 10.1002/pro.5560060408. Protein Sci. 1997. PMID: 9098891 Free PMC article.
-
The C9 methyl group of retinal interacts with glycine-121 in rhodopsin.Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13442-7. doi: 10.1073/pnas.94.25.13442. Proc Natl Acad Sci U S A. 1997. PMID: 9391044 Free PMC article.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
