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Spectroscopic characterization of reaction centers of the (M)Y210W mutant of the photosynthetic bacterium Rhodobacter sphaeroides

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Abstract

The tyrosine-(M)210 of the reaction center of Rhodobacter sphaeroides 2.4.1 has been changed to a tryptophan using site-directed mutagenesis. The reaction center of this mutant has been characterized by low-temperature absorption and fluorescence spectroscopy, time-resolved sub-picosecond spectroscopy, and magnetic resonance spectroscopy. The charge separation process showed bi-exponential kinetics at room temperature, with a main time constant of 36 ps and an additional fast time constant of 5.1 ps. Temperature dependent fluorescence measurements predict that the lifetime of P* becomes 4–5 times slower at cryogenic temperatures. From EPR and absorbance-detected magnetic resonance (ADMR, LD-ADMR) we conclude that the dimeric structure of P is not significantly changed upon mutation. In contrast, the interaction of the accessory bacteriochlorophyll BA with its environment appears to be altered, possibly because of a change in its position.

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Abbreviations

ADMR -:

absorbance-detected magnetic resonance

LDAO -:

N, N dimethyl dodecyl amine-N-oxide

RC -:

reaction center

LD-ADMR -:

linear-dichroic absorbance-detected magnetic resonance

P -:

primary donor

B -:

accessory bacteriochlorophyll

Ф -:

bacteriopheophytin

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Authors and Affiliations

  1. Department of Biophysics, Huygens Laboratory, Leiden University, P.O. Box 9504, 2300 RA, Leiden, The Netherlands

    Susana Shochat, Christof Francke, Peter Gast, Paula I. van Noort, Stephan C. M. Otte, Hans P. M. Schelvis, Jacobien Vrieze & Arnold J. Hoff

  2. Physik Department E 11, Technische Universität München, James Franck Str., 85748, Garching, Germany

    Thomas Arlt & Stefan Schmidt

  3. Department of Chemistry, Gorlaeus Laboratory, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands

    Erik Vijgenboom

  4. Institut für Medizinische Optik, Ludwig-Maximilians-Universität München, Barbarastr. 16, 80797, München, Germany

    Wolfgang Zinth

Authors
  1. Susana Shochat
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  2. Thomas Arlt
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  3. Christof Francke
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  4. Peter Gast
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  5. Paula I. van Noort
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  6. Stephan C. M. Otte
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  7. Hans P. M. Schelvis
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  8. Stefan Schmidt
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  9. Erik Vijgenboom
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  10. Jacobien Vrieze
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  11. Wolfgang Zinth
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  12. Arnold J. Hoff
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Shochat, S., Arlt, T., Francke, C. et al. Spectroscopic characterization of reaction centers of the (M)Y210W mutant of the photosynthetic bacterium Rhodobacter sphaeroides . Photosynth Res 40, 55–66 (1994). https://doi.org/10.1007/BF00019045

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  • DOI: https://doi.org/10.1007/BF00019045

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