Spectral and kinetic evidence for the existence of two forms of bathorhodopsin.

Einterz, C. M.; Lewis, James W.; Kliger, David S.

doi:10.1073/pnas.84.11.3699

Cited by 25 publications

(24 citation statements)

References 28 publications

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“…The broader half bandwidth of bathorhodopsin than that of rhodopsin may be explained by an existence of two kinds of bathorhodopsin, which was first reported by low temperature spectrophotometry (17,18) and then confirmed by nanosecond laser photolysis (19).…”

Section: Resultsmentioning

confidence: 91%

Absolute absorption spectra of batho- and photorhodopsins at room temperature. Picosecond laser photolysis of rhodopsin in polyacrylamide

Kandori¹,

Shichida²,

Yoshizawa³

1989

Biophysical Journal

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Picosecond laser photolysis of rhodopsin in 15% polyacrylamide gel was performed for estimating absolute absorption spectra of the primary intermediates of cattle rhodopsin (bathorhodopsin and photorhodopsin). Using a rhodopsin digitonin extract embedded in 15% polyacrylamide gel, a precise percentage of bleaching of rhodopsin after excitation of a picosecond laser pulse was measured. Using this value, the absolute absorption spectrum of bathorhodopsin was calculated from the spectral change before and 1 ns after the picosecond laser excitation (corresponding to the difference spectrum between rhodopsin and bathorhodopsin). The absorption spectrum of bathorhodopsin thus obtained displayed a lambda max at 535 nm, which was shorter than that at low temperature (543 nm) and a half band-width broader than that measured at low temperature. The oscillator strength of bathorhodopsin at room temperature was smaller than that at low temperature. The absolute absorption spectrum of photorhodopsin was also estimated from the difference spectrum measured at 15 ps after the excitation of rhodopsin (Shichida, Y., S. Matuoka, and T. Yoshizawa. 1984. Photobiochem. Photobiophys. 7:221-228), assuming a sequential conversion of photorhodopsin to bathorhodopsin. Its lambda max was located at approximately 570 nm, and the oscillator strength was smaller than those of rhodopsin and bathorhodopsin.

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Section: Resultsmentioning

confidence: 91%

Absolute absorption spectra of batho- and photorhodopsins at room temperature. Picosecond laser photolysis of rhodopsin in polyacrylamide

Kandori¹,

Shichida²,

Yoshizawa³

1989

Biophysical Journal

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“…Approximately 20 such groups were collected at each time delay, interspersed with the measurements at other delay times so that possible systematic errors, such as drifts in laser power, would not affect results. The results of each day were checked to see that the progressive red shift of the isosbestic point between difference spectra was observed as the delay time increased (12). If one of the averaged difference spectra was displaced by a probe lamp intensity fluctuation so that this trend was reversed, an offset was added to that trace to restore the trend.…”

Section: Sample Preparationmentioning

confidence: 99%

Absorbance Changes by Aromatic Amino Acid Side Chains in Early Rhodopsin Photointermediates

Lewis

Jäger

Kliger

1997

Photochem & Photobiology

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Absorbance changes were monitored from 250 to 650 nm during the first microsecond after photolysis of detergent suspensions of bovine rhodopsin at 20 degrees C. Global analysis of the resulting data produced difference spectra for bathorhodopsin, BSI and lumirhodopsin which give the change in absorbance of the aromatic amino acid side chains in these photointermediates relative to rhodopsin. These spectra show that the significant bleaching of absorbance near 280 nm, which has been seen previously for the lumirhodopsin, metarhodopsin I and metarhodopsin II intermediates, extends to times as early as bathorhodopsin. Because no corresponding absorbance increase is observed in the 250-275 nm region, the earliest bleaching of the 280 nm absorbance in rhodopsin is attributed to disruption of a hyperchromic interaction affecting Trp265. Partial decay of this 280 nm bleaching as bathorhodopsin converts to BSI takes place maximally near 290 nm, where Trp265 has been shown to absorb, and could be due to the ring of the retinylidene chromophore resuming a position at the BSI stage that reestablishes the hyperchromic interaction with Trp265. A subsequent change in the 250-300 nm region, which has no counterpart in the visible chromophore bands, indicates the possible presence of a protein-localized process as lumirhodopsin is formed.

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“…The optical density of the solutions (1-cm path length) used in the nanosecond experiments was typically 0.6 at the X.. of the pigment (461 nm in the case of cis-5,6-diH-ISORHO). Nanosecond photolysis measurements were performed as previously described (Einterz et al, 1987;Lewis et al, 1987) except that the actinic source was a Nd/YAG pumped dye laser which produced a 7-ns fwhm pulse of 477-nm light. Pulses with typical energies of 0.5 mJ were used to irradiate a sample area of 1 x 10 mm.…”

Section: Regeneration / Nanosecond Photolysismentioning

confidence: 99%

Photolysis intermediates of the artificial visual pigment cis-5,6-dihydro-isorhodopsin

Albeck

Friedman

Ottolenghi

et al. 1989

Biophysical Journal

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The photolysis intermediates of an artificial bovine rhodopsin pigment, cis-5,6-dihydro-isorhodopsin (cis-5,6,-diH-ISORHO, lambda max 461 nm), which contains a cis-5,6-dihydro-9-cis-retinal chromophore, are investigated by room temperature, nanosecond laser photolysis, and low temperature irradiation studies. The observations are discussed both in terms of low temperature experiments of Yoshizawa and co-workers on trans-5,6-diH-ISORHO (Yoshizawa, T., Y. Shichida, and S. Matuoka. 1984. Vision Res. 24: 1455-1463), and in relation to the photolysis intermediates of native bovine rhodopsin (RHO). It is suggested that in 5,6-diH-ISORHO, a primary bathorhodopsin intermediate analogous to the bathorhodopsin intermediate (BATHO) of the native pigment, rapidly converts to a blue-shifted intermediate (BSI, lambda max 430 nm) which is not observed after photolysis of native rhodopsin. The analogs from lumirhodopsin (LUMI) to meta-II rhodopsin (META-II) are generated subsequent to BSI, similar to their generation from BATHO in the native pigment. It is proposed that the retinal chromophore in the bathorhodopsin stage of 5,6-diH-ISORHO is relieved of strain induced by the primary cis to trans isomerization by undergoing a geometrical rearrangement of the retinal. Such a rearrangement, which leads to BSI, would not take place so rapidly in the native pigment due to ring-protein interactions. In the native pigment, the strain in BATHO would be relieved only on a longer time scale, via a process with a rate determined by protein relaxation.

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Spectral and kinetic evidence for the existence of two forms of bathorhodopsin.

Cited by 25 publications

References 28 publications

Absolute absorption spectra of batho- and photorhodopsins at room temperature. Picosecond laser photolysis of rhodopsin in polyacrylamide

Absolute absorption spectra of batho- and photorhodopsins at room temperature. Picosecond laser photolysis of rhodopsin in polyacrylamide

Absorbance Changes by Aromatic Amino Acid Side Chains in Early Rhodopsin Photointermediates

Photolysis intermediates of the artificial visual pigment cis-5,6-dihydro-isorhodopsin

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