Proton uptake of Rhodobacter capsulatus reaction center mutants modified in the primary quinone environment

Julia Tandori, Jaroslava Miksovska, Marielle Valerio-Lepiniec, Marianne Schiffer, Peter Maróti, Deborah K. Hanson, Pierre Sebban

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Flash-induced absorbance spectroscopy was used to analyze the proton uptake and electron transfer properties of photosynthetic reaction centers (RC) of Rhodobacter capsulatus that have been genetically modified near the primary quinone electron acceptor (QA). M246AIa and M247AIa, which are symmetry-related to the positions of two acidic groups, L212Glu and L213Asp, in the secondary quinone electron acceptor (QB) protein environment, have been mutated to Glu and Asp, respectively. The pH dependence of the stoichiometry of proton uptake upon formation of the P+Q A- (H+/P+QA-) and (H+/QA-) (P is the primary electron donor, a noncovalently linked bacteriochlorophyll dimer) states have been measured in the M246AIa → Glu and the M247AIa → Asp mutant RC, in the M246AIa-M247AIa → Glu-Asp double mutant and in the wild type (WT). Our results show that the introduction of an acidic group (Glu or Asp) in the Q A protein region induces notable additional proton uptake over a large pH region (∼6-9), which reflects a delocalized response of the protein to the formation of QA- This may indicate the existence of a widely spread proton reservoir in the cytoplasmic region of the protein. Interestingly, the pH titration curves of the proton release caused by the formation of P+ (H+/P+: difference between H+/P+QA- and H+/PQ A- curves) are nearly superimposable in the WT and the M246AIa → Glu mutant RC, but substantial additional proton release is detected between pH 7 and 9 in the M247AIa → Asp mutant RC. This effect can be accounted for by an increased proton release by the P+ environment in the M247AIa → Asp mutant. The M247AIa → Asp mutation reveals the existence of an energetic and conformational coupling between donor and acceptor sides of the RC at a distance of nearly 30Å.

Original languageEnglish
Pages (from-to)126-133
Number of pages8
JournalPhotochemistry and photobiology
Volume75
Issue number2
DOIs
Publication statusPublished - Feb 1 2002

ASJC Scopus subject areas

  • Biochemistry
  • Physical and Theoretical Chemistry

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