The identification of Lys<sub>216</sub> as the retinal binding residue in bacteriorhodopsin

Mullen, Eric; Johnson, Alan; Akhtar, Muhammad

doi:10.1016/0014-5793(81)81116-7

Cited by 58 publications

(18 citation statements)

References 11 publications

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“…This concept has its origins in physical studies on bacteriorhodopsin. The all-trans-retinal chromophore of bacteriorhodopsin is attached via a Schiff base to Lys216, located in the seventh transmembrane helix of the protein (Bayley et al, 1981;Mullen et al, 1981), and is contained within a pocket formed by the transmembrane helices (Henderson et al, 1990;see Fig. 1).…”

Section: The Search For Features Common To G Protein-coupled Receptormentioning

confidence: 99%

In vitro mutagenesis and the search for structure-function relationships among G protein-coupled receptors

Savarese

Fraser

1992

Biochemical Journal

462

236

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Section: The Search For Features Common To G Protein-coupled Receptormentioning

confidence: 99%

In vitro mutagenesis and the search for structure-function relationships among G protein-coupled receptors

Savarese

Fraser

1992

Biochemical Journal

462

236

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“…In either rhodopsin (Bownds and Wald, 1965) or B R (Bayley et al, 1981;Lemke and Oesterhelt, 1981;Mullen et al, 1981) the retinyl chromophores are known to be attached to the eaniino group of a lysine residue via a protonated Schiff base linkage (Mathies et al, 1976;Aton et al, 1977). A possible role of the butyl side chain of the appended lysine has been considered.…”

Section: Introductionmentioning

confidence: 99%

BUTYL CONFORMATIONAL REORGANIZATION AS A POSSIBLE EXPLANATION FOR THE LONGITUDINAL FLEXIBILITY OF THE BINDING SITE OF BACTERIORHODOPSIN. THE AZULENE and C‐22 RETINOID ANALOGS*

Liu

et al. 1991

Photochem & Photobiology

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The UV-VIS absorption data of four bacteriorhodopsin (BR) analogs formed from azulene-retinals of varying polyene chain length show that the one-bond-shortened to one-bond-lengthened analogs possess comparable opsin shift values to that of BR. A two-bond-shortened analog exhibited a much smaller opsin shift. These data, combined with those reported for the C-22 retinal analog (Tokunaga et al., 1977, Biophys. J. 19, 191-198) were analyzed by molecular modelling and computer graphics in terms of a model where conformational flexibility of the appended butyl is the controlling factor in determining ease of pigment formation and protein/substrate interaction.

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“…This peak is normally due to fragment CNBr-1 (19). Little, if any, retinyl absorbance comigrates with the only tryptophan-containing fragment, CNBr-2 (amino acids [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20], identified by the characteristic shoulder near 290 nm (Fig. 2).…”

Section: Resultsmentioning

confidence: 99%

Retinal migration during dark reduction of bacteriorhodopsin

Wolber

Stoeckenius²

1984

Proc. Natl. Acad. Sci. U.S.A.

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When the retinal Schiff base in chymotryptically cleaved bacteriorhodopsin is reduced to a secondary retinylamine by prolonged exposure to 10% (wt/vol) sodium cyanoborohydride, at pH 10, in the absence of light, =45% of the retinal is found linked to , and the remainder is scattered over various sites on the large chymotryptic fragment, including the physiological site at Lys-216. The retinal-binding site is destroyed or blocked by the reduction conditions, but the bacteriorhodopsin lattice remains intact. The results demonstrate that artifactual linkage to Lys-40/41 is possible under special conditions. Under these conditions, the e-amino groups of Lys-40/41 show an enhanced ability to form retinylidene linkages with the retinal released by the physiological linkage site at Lys-216, due to some combination of close proximity to the normal linkage site, and increased reactivity with respect to other lysine Eamino groups. The results are of interest for the characterization of the two newly discovered rhodopsin-like proteins, halorhodopsin and slow rhodopsin.The determination of the primary protein (1, 2) and structural gene (3) sequences of bacteriorhodopsin (bR), the lightdriven proton pump from the purple membrane (pm) of Halobacterium halobium, has led to a reinvestigation of the retinal-binding site by chemical reduction and peptide analysis. This resulted in a reassignment for the binding site from the E-amino group of to the E-amino group of Lys-216 (5-9). Additional chemical modification (10) and resonance Raman (11) studies have demonstrated conclusively that the binding site does not change during the photoreaction cycle as originally postulated by Ovchinnikov's group (12). The reason for the original misassignment of the binding site by Bridgen and Walker is obvious in retrospect (7). However, the same arguments cannot explain the more recent observations of reductive retinal linkage to Lys-40/41 (7,9,12). Lemke and Oesterhelt (6) consider these to be in error due to a previously unnoticed proteolytic cleavage of bR by NaBH4. We show here that this explanation does not hold, at least for our results, and that under some conditions of reduction, two-thirds of the retinal are indeed found bound to Lys-40/41.More important, we have previously argued that while binding to Lys-40/41 is a preparation artifact, it might still be used to obtain important information about the tertiary structure of bR. We have, therefore, repeated and extended our earlier experiments. Unfortunately, the new observations considerably weaken the structural argument, but they demonstrate an unusually high reactivity of Lys-40/41 for Schiff base formation and/or reduction and are of considerable interest for the interpretation of experiments trying to identify the two additional retinal pigments recently discovered in H. halobium (13)(14)(15) Repenerated, Chymotryptically Cut, [3H]Retinal-Labeled pm ( H-RG-CT-pm). RG-CT-pm was prepared essentially as described (7, 17), except that hydroxylamine-bleached pm was cut at a c...

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The identification of Lys₂₁₆ as the retinal binding residue in bacteriorhodopsin

Cited by 58 publications

References 11 publications

In vitro mutagenesis and the search for structure-function relationships among G protein-coupled receptors

In vitro mutagenesis and the search for structure-function relationships among G protein-coupled receptors

BUTYL CONFORMATIONAL REORGANIZATION AS A POSSIBLE EXPLANATION FOR THE LONGITUDINAL FLEXIBILITY OF THE BINDING SITE OF BACTERIORHODOPSIN. THE AZULENE and C‐22 RETINOID ANALOGS*

Retinal migration during dark reduction of bacteriorhodopsin

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The identification of Lys216 as the retinal binding residue in bacteriorhodopsin

Cited by 58 publications

References 11 publications

In vitro mutagenesis and the search for structure-function relationships among G protein-coupled receptors

In vitro mutagenesis and the search for structure-function relationships among G protein-coupled receptors

BUTYL CONFORMATIONAL REORGANIZATION AS A POSSIBLE EXPLANATION FOR THE LONGITUDINAL FLEXIBILITY OF THE BINDING SITE OF BACTERIORHODOPSIN. THE AZULENE and C‐22 RETINOID ANALOGS*

Retinal migration during dark reduction of bacteriorhodopsin

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The identification of Lys₂₁₆ as the retinal binding residue in bacteriorhodopsin