### Abstract

The kinetics of electron transfer from cytochrome c_{2} to the primary donor (P) of the reaction center from the photosynthetic purple bacterium Rhodobacter sphaeroides have been investigated by time-resolved absorption spectroscopy. Rereduction of P^{+} induced by a laser pulse has been measured at temperatures from 300 K to 220 K in a series of specifically mutated reaction centers characterized by altered midpoint redox potentials of P^{+}/P varying from 410 mV to 765 mV (as compared to 505 mV for wild type). Rate constants for first-order electron donation within preformed reaction center- cytochrome c_{2} complexes and for the bimolecular oxidation of free cytochrome c_{2} have been obtained by multiexponential deconvolution of the kinetics. At all temperatures the rate of the fastest intracomplex electron transfer increases by more than two orders of magnitude as the driving force -ΔG°is varied over a range of 350 meV. The temperature and ΔG°dependences of the rate constant fit the Marcus equation well. Global analysis yields a reorganization energy λ = 0.96 ± 0.07 eV and a set of electronic matrix elements, specific for each mutant, ranging from 1.2 10^{-4} eV to 2.5 10^{-4} eV. Analysis in terms of the Jortner equation indicates that the best fit is obtained in the classical limit and restricts the range of coupled vibrational modes to frequencies lower than ~200 cm^{-1}. An additional slower kinetic component of P^{+} reduction, attributed to electron transfer from cyt c_{2} docked in a nonoptimal configuration of the complex, displays a Marcus type dependence of the rate constant upon ΔG°, characterized by a similar value of λ (0.8 ± 0.1 eV) and by an average electronic matrix element smaller by more than one order of magnitude. In all of the mutants, as the temperature is decreased below 260 K, both intracomplex reactions are abruptly inhibited, their rate being negligible at 220 K. The free energy dependence of the second-order rate constant for oxidation of cyt c_{2} in solution suggests that the collisional reaction is partially diffusion controlled, reaching the diffusion limit at exothermicities between 150 and 250 meV over the temperature range investigated.

Original language | English (US) |
---|---|

Pages (from-to) | 3226-3240 |

Number of pages | 15 |

Journal | Biophysical Journal |

Volume | 74 |

Issue number | 6 |

State | Published - Jun 1998 |

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### ASJC Scopus subject areas

- Biophysics

### Cite this

_{2}to the photosynthetic reaction center of the purple bacterium Rhodobacter sphaeroides.

*Biophysical Journal*,

*74*(6), 3226-3240.

**Effects of temperature and ΔG°on electron transfer from cytochrome c _{2} to the photosynthetic reaction center of the purple bacterium Rhodobacter sphaeroides.** / Venturoli, G.; Drepper, F.; Williams, Joann; Allen, James; Lin, X.; Mathis, P.

Research output: Contribution to journal › Article

_{2}to the photosynthetic reaction center of the purple bacterium Rhodobacter sphaeroides',

*Biophysical Journal*, vol. 74, no. 6, pp. 3226-3240.

_{2}to the photosynthetic reaction center of the purple bacterium Rhodobacter sphaeroides. Biophysical Journal. 1998 Jun;74(6):3226-3240.

}

TY - JOUR

T1 - Effects of temperature and ΔG°on electron transfer from cytochrome c2 to the photosynthetic reaction center of the purple bacterium Rhodobacter sphaeroides

AU - Venturoli, G.

AU - Drepper, F.

AU - Williams, Joann

AU - Allen, James

AU - Lin, X.

AU - Mathis, P.

PY - 1998/6

Y1 - 1998/6

N2 - The kinetics of electron transfer from cytochrome c2 to the primary donor (P) of the reaction center from the photosynthetic purple bacterium Rhodobacter sphaeroides have been investigated by time-resolved absorption spectroscopy. Rereduction of P+ induced by a laser pulse has been measured at temperatures from 300 K to 220 K in a series of specifically mutated reaction centers characterized by altered midpoint redox potentials of P+/P varying from 410 mV to 765 mV (as compared to 505 mV for wild type). Rate constants for first-order electron donation within preformed reaction center- cytochrome c2 complexes and for the bimolecular oxidation of free cytochrome c2 have been obtained by multiexponential deconvolution of the kinetics. At all temperatures the rate of the fastest intracomplex electron transfer increases by more than two orders of magnitude as the driving force -ΔG°is varied over a range of 350 meV. The temperature and ΔG°dependences of the rate constant fit the Marcus equation well. Global analysis yields a reorganization energy λ = 0.96 ± 0.07 eV and a set of electronic matrix elements, specific for each mutant, ranging from 1.2 10-4 eV to 2.5 10-4 eV. Analysis in terms of the Jortner equation indicates that the best fit is obtained in the classical limit and restricts the range of coupled vibrational modes to frequencies lower than ~200 cm-1. An additional slower kinetic component of P+ reduction, attributed to electron transfer from cyt c2 docked in a nonoptimal configuration of the complex, displays a Marcus type dependence of the rate constant upon ΔG°, characterized by a similar value of λ (0.8 ± 0.1 eV) and by an average electronic matrix element smaller by more than one order of magnitude. In all of the mutants, as the temperature is decreased below 260 K, both intracomplex reactions are abruptly inhibited, their rate being negligible at 220 K. The free energy dependence of the second-order rate constant for oxidation of cyt c2 in solution suggests that the collisional reaction is partially diffusion controlled, reaching the diffusion limit at exothermicities between 150 and 250 meV over the temperature range investigated.

AB - The kinetics of electron transfer from cytochrome c2 to the primary donor (P) of the reaction center from the photosynthetic purple bacterium Rhodobacter sphaeroides have been investigated by time-resolved absorption spectroscopy. Rereduction of P+ induced by a laser pulse has been measured at temperatures from 300 K to 220 K in a series of specifically mutated reaction centers characterized by altered midpoint redox potentials of P+/P varying from 410 mV to 765 mV (as compared to 505 mV for wild type). Rate constants for first-order electron donation within preformed reaction center- cytochrome c2 complexes and for the bimolecular oxidation of free cytochrome c2 have been obtained by multiexponential deconvolution of the kinetics. At all temperatures the rate of the fastest intracomplex electron transfer increases by more than two orders of magnitude as the driving force -ΔG°is varied over a range of 350 meV. The temperature and ΔG°dependences of the rate constant fit the Marcus equation well. Global analysis yields a reorganization energy λ = 0.96 ± 0.07 eV and a set of electronic matrix elements, specific for each mutant, ranging from 1.2 10-4 eV to 2.5 10-4 eV. Analysis in terms of the Jortner equation indicates that the best fit is obtained in the classical limit and restricts the range of coupled vibrational modes to frequencies lower than ~200 cm-1. An additional slower kinetic component of P+ reduction, attributed to electron transfer from cyt c2 docked in a nonoptimal configuration of the complex, displays a Marcus type dependence of the rate constant upon ΔG°, characterized by a similar value of λ (0.8 ± 0.1 eV) and by an average electronic matrix element smaller by more than one order of magnitude. In all of the mutants, as the temperature is decreased below 260 K, both intracomplex reactions are abruptly inhibited, their rate being negligible at 220 K. The free energy dependence of the second-order rate constant for oxidation of cyt c2 in solution suggests that the collisional reaction is partially diffusion controlled, reaching the diffusion limit at exothermicities between 150 and 250 meV over the temperature range investigated.

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M3 - Article

C2 - 9635776

AN - SCOPUS:0031595249

VL - 74

SP - 3226

EP - 3240

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 6

ER -