Modification of Ovine Opsin with the Photosensitive Hydrophobic Probe 1-azido-4-[125I]iodobenzene. Labelling of the Chromophore-attachment Domain

Overview

Journal Biochem J

Specialty Biochemistry

Date 1986 Mar 1

PMID 2941011

Citations 9

Authors

M D Davison

J B Findlay

Affiliations

Soon will be listed here.

Abstract

The hydrophobic photosensitive probe 1-azido-4-[125I]iodobenzene (AIB) partitioned preferentially into photoreceptor disc membranes and, upon u.v. irradiation, became covalently bound to opsin and phospholipid. The labelling of both protein and phospholipid was linearly related to AIB concentration. The amount of probe incorporated into protein was not significantly different when membranes were irradiated at -100 degrees, 4 degrees or 25 degrees C, but irreversible aggregation of monomeric opsin was dramatically reduced by performing the photolysis at -100 degrees C. Labelling of opsin after irradiation at -100 degrees or 4 degrees was not significantly reduced by the presence of lysine in the aqueous buffer, indicating that significant amounts of reactive species did not enter the aqueous phase. The incorporation into phospholipid, unlike that into opsin, decreased as the temperature of irradiation increased. Some labelling of opsin occurred on incubation with pre-photoactivated AIB, indicating that reaction may also occur with reactive species of longer lifetimes than the nitrene. Proteolysis of labelled opsin with Staphylococcus aureus V8 proteinase yielded two radiolabelled membrane-bound fragments. The location of the modified sites (cysteine, tryptophan, tyrosine, lysine and histidine residues: all nucleophiles) in the smaller fragment was entirely consistent with putative models for the protein derived from other studies.

Citing Articles

Homology modelling and spectroscopy, a never-ending love story.

Venselaar H, Joosten R, Vroling B, Baakman C, Hekkelman M, Krieger E Eur Biophys J. 2009; 39(4):551-63.

PMID: 19718498 PMC: 2841279. DOI: 10.1007/s00249-009-0531-0.

Positioning of proteins in membranes: a computational approach.

Lomize A, Pogozheva I, Lomize M, Mosberg H Protein Sci. 2006; 15(6):1318-33.

PMID: 16731967 PMC: 2242528. DOI: 10.1110/ps.062126106.

The transmembrane 7-alpha-bundle of rhodopsin: distance geometry calculations with hydrogen bonding constraints.

Pogozheva I, Lomize A, Mosberg H Biophys J. 1997; 72(5):1963-85.

PMID: 9129801 PMC: 1184393. DOI: 10.1016/S0006-3495(97)78842-8.

Sequencing and identification of a cDNA clone for tomato polygalacturonase.

Grierson D, Tucker G, Keen J, Ray J, Bird C, Schuch W Nucleic Acids Res. 1986; 14(21):8595-603.

PMID: 3786135 PMC: 311879. DOI: 10.1093/nar/14.21.8595.

On the disulphide bonds of rhodopsins.

Al-Saleh S, Gore M, Akhtar M Biochem J. 1987; 246(1):131-7.

PMID: 3675552 PMC: 1148249. DOI: 10.1042/bj2460131.

References

Pober J, Stryer L . Letter to the editor: Light dissociates enzymatically-cleaved rhodopsin into two different fragments. J Mol Biol. 1975; 95(3):477-81. DOI: 10.1016/0022-2836(75)90204-1. View

Findlay J, Barclay P, Brett M, Davison M, Pappin D, Thompson P . The structure of mammalian rod opsins. Vision Res. 1984; 24(11):1501-8. DOI: 10.1016/0042-6989(84)90312-2. View

Karlish S, Jorgensen P, Gitler C . Identification of a membrane-embedded segment of the large polypeptide chain of (Na+, K+)ATPase. Nature. 1977; 269(5630):715-7. DOI: 10.1038/269715a0. View

Kahane I, Gitler C . Red cell membrane glycophorin labeling from within the lipid bilayer. Science. 1978; 201(4353):351-2. DOI: 10.1126/science.663661. View

Bayley H, Knowles J . Photogenerated reagents for membrane labeling. 1. Phenylnitrene formed within the lipid bilayer. Biochemistry. 1978; 17(12):2414-9. DOI: 10.1021/bi00605a025. View

Bayley H, Knowles J . Photogenerated reagents for membrane labeling. 2. Phenylcarbene and adamantylidene formed within the lipid bilayer. Biochemistry. 1978; 17(12):2420-3. DOI: 10.1021/bi00605a026. View

Brett M, Findlay J . Investigation of the organization of rhodopsin in the sheep photoreceptor membrane by using cross-linking reagents. Biochem J. 1979; 177(1):215-23. PMC: 1186359. DOI: 10.1042/bj1770215. View

Steffens G, Buse G . Studies on cytochrome c oxidase, IV[1--3]. Primary structure and function of subunit II. Hoppe Seylers Z Physiol Chem. 1979; 360(4):613-9. View

Gupta C, Radhakrishnan R, Gerber G, Olsen W, Quay S, Khorana H . Intermolecular crosslinking of fatty acyl chains in phospholipids: use of photoactivable carbene precursors. Proc Natl Acad Sci U S A. 1979; 76(6):2595-9. PMC: 383654. DOI: 10.1073/pnas.76.6.2595. View

10.

GOLDMAN D, Pober J, White J, Bayley H . Selective labelling of the hydrophobic segments of intrinsic membrane proteins with a lipophilic photogenerated carbene. Nature. 1979; 280(5725):841-3. DOI: 10.1038/280841a0. View

11.

Wells E, Findlay J . Labelling of the intramembranous region of the major sialoglycoprotein of human erythrocytes with a photosensitive hydrophobic probe. Biochem J. 1979; 179(2):265-72. PMC: 1186623. DOI: 10.1042/bj1790265. View

12.

Hoppe J, Friedl P, Schairer H, Sebald W, von Meyenburg K, Jorgensen B . The topology of the proton translocating F0 component of the ATP synthase from E. coli K12: studies with proteases. EMBO J. 1983; 2(1):105-10. PMC: 555095. DOI: 10.1002/j.1460-2075.1983.tb01389.x. View

13.

HELMKAMP R, Sears D . A label for the red cell membrane: diazotized diiodosulfanilic acid. Int J Appl Radiat Isot. 1970; 21(11):683-5. DOI: 10.1016/0020-708x(70)90127-4. View

14.

Spatz L, STRITTMATTER P . A form of cytochrome b5 that contains an additional hydrophobic sequence of 40 amino acid residues. Proc Natl Acad Sci U S A. 1971; 68(5):1042-6. PMC: 389109. DOI: 10.1073/pnas.68.5.1042. View

15.

Horn M, Laursen R . Solid-phase edman degradation: attachment of carboxyl-terminal homoserine peptides to an insoluble resin. FEBS Lett. 1973; 36(3):285-8. DOI: 10.1016/0014-5793(73)80392-8. View

16.

Daemen F . Vertebrate rod outer segment membranes. Biochim Biophys Acta. 1973; 300(3):255-88. DOI: 10.1016/0304-4157(73)90006-3. View

17.

Laemmli U, Favre M . Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973; 80(4):575-99. DOI: 10.1016/0022-2836(73)90198-8. View

18.

Klip A, Gitler C . Photoactive covalent labeling of membrane components from within the lipid core. Biochem Biophys Res Commun. 1974; 60(3):1155-62. DOI: 10.1016/0006-291x(74)90433-1. View

19.

Eytan G, Schatz G . Cytochrome c oxidase from bakers' yeast. V. Arrangement of the subunits in the isolated and membrane-bound enzyme. J Biol Chem. 1975; 250(2):767-74. View

20.

Wells E, Findlay J . Use of photosensitive hydrophobic probes to label the membrane of the human erythrocyte. Biochem J. 1979; 179(2):257-64. PMC: 1186622. DOI: 10.1042/bj1790257. View