Femtosecond Dynamics of an Excited Primary Electron Donor in Reaction Centers of the Purple Bacterium Rhodobacter sphaeroides

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Abstract

Femtosecond transient absorption spectroscopy spectroscopy was used to study the dynamics of the excited primary electron donor in the reaction centers of the purple bacterium Rhodobacter sphaeroides. Using global analysis and the interval method, a correlation was found between the vibrational coherence damping of the excited primary electron donor and the lifetime of the charge-separated state P+BA–, indicating the reversibility of electron transfer to the primary electron acceptor, the BA molecule. In the reaction centers, signs of superposition of two electronic states of P were found for a delay time of less than 200 fs. It is suggested that the admixture value of charge transfer state PA+PB– with the excited primary electron donor P* is about 24%. The results obtained are discussed in terms of the two-step electron transfer mechanism.

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A. M. Khristin

Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences

Email: rgreen1@rambler.ru
Russian Federation, Pushchino

T. Yu. Fufina

Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences

Email: rgreen1@rambler.ru
Russian Federation, Pushchino

R. A. Khatypov

Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences

Author for correspondence.
Email: rgreen1@rambler.ru
Russian Federation, Pushchino

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3. Fig. 1. Differential absorption spectra of RCs of Rba. Sphaeroides (a) at time delays of 40 fs (curve 1), 180 fs (curve 2), 300 fs (curve 3), 1.5 ps (curve 4) and 210 ps (curve 5) measured at 293 K. b - Double-differential absorption spectra obtained by subtracting spectra at time delays of 40 fs minus 210 ps (curve 1), 1.5 ps minus 210 ps (curve 2), 180 fs minus 1.5 ps (curve 3) and 300 fs minus 1.5 ps (curve 4); the dashed line shows the ∆A spectrum measured at a time delay of 210 ps

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4. Fig. 2. Kinetic curves of absorbance changes at 870 nm (a), at 940 nm (b) and at 915 nm (c) in the RC of Rba. sphaeroides obtained by the interval method at 293 K. The contribution of stimulated emission is highlighted in grey in the figure. The solid line shows a single exponent approximation of the kinetics at 915 nm. The dotted line shows positive changes due to the contribution of the fading of the absorption band of the primary donor P. The inset shows the kinetic curve at 940 nm on a time scale of -0.5 to 10 ps

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5. Fig. 3. Stimulated emission kinetics at 940 nm (open symbols) and absorption band kinetics of the anion radical BA- at 1025 nm (closed symbols) measured at 293 K (a) and at 100 K (b) in the RC of Rba. sphaeroides. The amplitude of the first peak of stimulated emission is normalised to the maximum rise of the absorption band at 1025 nm. The double-sided arrow shows the time delay at which the complete attenuation of emission coincides with the complete increase in absorption of BA-

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6. Fig. 4. Exponential decay-associated difference spectra (decay-associated difference spectra, DADS) of RC absorption spectra of Rba. sphaeroides measured at physiological temperature (a) and at 100 K (b)

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7. Fig. 5. Differential absorption spectra of wild-type RC Rba. sphaeroides at 60 fs (curve 1) and 170 fs (curve 2) time delays (a) and RC of the triple mutant at 80 fs (curve 1) and 380 fs (curve 2) time delays (b), measured at 100 K. The insets (panels a and b) show the region of the P* absorption band at 810 nm. The magnitude of the positive dispersion of the probing pulse in the wavelength range 810-940 nm is ~100 fs

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8. Fig. 6. Twice differential absorption spectra of the RC mutant M160LH + L131LH + M197FH of Rba. sphaeroides obtained by subtracting the differential spectrum measured at a time delay of 220 fs from the differential spectra measured at time delays of 260 fs (curve 1), 300 fs (curve 2), and 340 fs (curve 3) at 100 K temperature. The arrow shows the wavelength of the maximum of the absorption band P

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9. Fig. 7. Kinetic curves of the absorption band at 810 nm in the RCs of wild-type Rba. sphaeroides (curve 1) and the M160LH + L131LH + M197FH mutant (curve 2) at 100 K. The dotted line shows the amplitude of the absorption band at the interaction of the excitation and probing pulses, and the dashed line shows the increase of the absorption band in the RC of the mutant at time delays longer than the duration of the hardware function (double-sided arrow)

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