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• W2067236472 abstract "Aqueous suspensions of dark-adapted bacteriorhodopsin (bR560) in the purple membrane of Halobacterium halobium are exposed to rapid jumps to high pH. Optical and resonance Raman measurements are carried out by using flow and stationary methods. Above pH congruent to 11.5 bR560 starts to be reversibly converted to a species absorbing at 460 nm (bR460) characterized by an unprotonated Schiff base chromophore. Above pH congruent to 13.0 bleaching takes place, first reversibly and subsequently irreversibly, to a species absorbing around 365 nm (bR365). This process competes with the formation of bR460. The pKa corresponding to the equilibrium (equation in text) is determined as 13.3 +/- 0.3. The value of the corresponding association rate constant determined from the reverse jumps (from pH 12.67 to pH 10 and 9.2) is ka = (3.5 +/- 0.5) X 10(11) M-1 s-1. Thus, starting with bR at pH 12.67 the reprotonation process is diffusion controlled as observed for homogeneous acid-base equilibria. The observed rate of dissociation when jumping from pH 6.5 to 12-13 is slower than that predicted by including the equilibrium (equation in text) The results imply that the Schiff base is titratable in the dark, but its accessibility to external OH- ions is limited. The limitations in the significance of the apparent value of pKa = 13.3 observed for the Schiff base titration are discussed in light of possible alterations in the structure of bR resulting from the parallel titration of other protein groups. It is suggested that a light-induced pKa change of at least nine units takes place during the photocycle of light-adapted bR." @default.
• W2067236472 created "2016-06-24" @default.
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• W2067236472 date "1982-09-28" @default.
• W2067236472 modified "2023-10-18" @default.
• W2067236472 title "Acid-base equilibrium of the Schiff base in bacteriorhodopsin" @default.
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• W2067236472 doi "https://doi.org/10.1021/bi00263a019" @default.
• W2067236472 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/7138840" @default.
• W2067236472 hasPublicationYear "1982" @default.
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