Realtime kinetics of the light driven steps of photosynthetic water oxidation in living organisms by “stroboscopic” fluorometry

Colin Gates, Gennady Ananyev, G. Charles Dismukes

Research output: Contribution to journalArticle

Abstract

We develop a rapid “stroboscopic” fluorescence induction method, using the fast repetition rate fluorometry (FRRF) technique, to measure changes in the quantum yield of light emission from chlorophyll in oxygenic photosynthesis arising from competition with primary photochemical charge separation (P680* ➔ P680 +QA ). This method determines the transit times of electrons that pass through PSII during the successive steps in the catalytic cycle of water oxidation/O2 formation (S states) and plastoquinone reduction in any oxygenic phototroph (in vivo or in vitro). We report the first measurements from intact living cells, illustrated by a eukaryotic alga (Nannochloropsis oceanica). We demonstrate that S state transition times depend strongly on the redox state of the PSII acceptor side, at both QB and the plastoquinone pool which serve as the major locus of regulation of PSII electron flux. We provide evidence for a kinetic intermediate S3′ state (lifetime 220 μs) following formation of S3 and prior to the release of O2. We compare the FRRF-detected kinetics to other previous spectroscopic methods (optical absorbance, EPR, and XES) that are applicable only to in vitro samples.

Original languageEnglish
Article number148212
JournalBiochimica et Biophysica Acta - Bioenergetics
Volume1861
Issue number8
DOIs
Publication statusPublished - Aug 1 2020

Keywords

  • Fluorometry
  • P680
  • Photosystem II
  • S-states
  • Transition times
  • Water oxidizing complex

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

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