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Difference Spectra Of Late Intermediates Of The Bacteriorhodopsin Photocycle

Authors: R. Govindjee; Zs. Dancshazy; T. G. Ebrey;

Difference Spectra Of Late Intermediates Of The Bacteriorhodopsin Photocycle

Abstract

The flash-induced difference absorbance spectra of bacteriorhodopsin (BR) were measured at various times after an actinic flash using a diode array spectrophotometer (300-700 nm). Difference spectra for three late bacteriorhodopsin photocycle intermediates Mfast (Mf), mslow (Ms) and R are reported. The main distinguishing features of the 3 difference spectra at pH = 10.5 and 5 °C are as follows: Mf ΔAmax = 412 nm, a shoulder at 436 nm, no absorbance change at 350 nm, ΔAmin = 565 nm, ΔA412/ ΔA565 = 0.8-0.9. Ms: ΔAmax = 412 nm, a shoulder at 386 nm, ΔAmin = 570-575 nm, ΔA.412/ ΔA575 = 0.6. R: ΔAmax = 336 and 350 nm (double peak), minor peaks at 386 and 412 nms,ΔAmin = 585-590 nm; ΔA350/ ΔA585 = 0.2. The t1/2 of Mf, Ms and R and the relative weights of BR570 recovered with these rates are: 1 sec (50%), 3-5 sec (25%) and 35 sec (25%) respectively. These spectral features can also be seen at pH = 7, -16 °C, and at pH = 9-10.5, 20 °C. Based on some assumptions, the absorption maximum of R was calculated to be at ca. 550 nm. The extinction coefficient of R is approximately 70% that of light-adapted BR. We suggest that: 1) Mf decays into R and R decays to BR, and 2) there are two types of BR with independent photocycles; Ms and probably 0 are in the photocycle near neutral pH, and Mf and R in a photocycle predominant at higher pH.

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
4
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