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Production of reactive oxygen species in decoupled, Ca2+-depleted PSII and their use in assigning a function to chloride on both sides of PSII

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Abstract

Extraction of Ca2+ from the oxygen-evolving complex of photosystem II (PSII) in the absence of a chelator inhibits O2 evolution without significant inhibition of the light-dependent reduction of the exogenous electron acceptor, 2,6-dichlorophenolindophenol (DCPIP) on the reducing side of PSII. The phenomenon is known as “the decoupling effect” (Semin et al. Photosynth Res 98:235–249, 2008). Extraction of Cl− from Ca2+-depleted membranes (PSII[–Ca]) suppresses the reduction of DCPIP. In the current study we investigated the nature of the oxidized substrate and the nature of the product(s) of the substrate oxidation. After elimination of all other possible donors, water was identified as the substrate. Generation of reactive oxygen species HO, H2O2, and O ·−2 , as possible products of water oxidation in PSII(–Ca) membranes was examined. During the investigation of O ·−2 production in PSII(–Ca) samples, we found that (i) O ·−2 is formed on the acceptor side of PSII due to the reduction of O2; (ii) depletion of Cl− does not inhibit water oxidation, but (iii) Cl− depletion does decrease the efficiency of the reduction of exogenous electron acceptors. In the absence of Cl− under aerobic conditions, electron transport is diverted from reducing exogenous acceptors to reducing O2, thereby increasing the rate of O ·−2 generation. From these observations we conclude that the product of water oxidation is H2O2 and that Cl− anions are not involved in the oxidation of water to H2O2 in decoupled PSII(–Ca) membranes. These results also indicate that Cl− anions are not directly involved in water oxidation by the Mn cluster in the native PSII membranes, but possibly provide access for H2O molecules to the Mn4CaO5 cluster and/or facilitate the release of H+ ions into the lumenal space.

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Abbreviations

Buffer A(–Cl):

Buffer A without Cl−

Chl:

Chlorophyll

DCPIP:

2,6-Dichlorophenol indophenol

DCMU:

3-(3,4-dichlorophenyl)-1,1-dimethylurea

Decoupled PSII(–Ca):

Ca2+-depleted PSII that are inhibited in O2 evolution without significant inhibition of the light-dependent reduction of an exogenous electron acceptor

DMAB:

3-(Dimethylamino) benzoic acid

DMBQ:

2,6-Dimethyl-p-benzoquinone

DPC:

1,5-Diphenycarbazide

HA site:

High-affinity Mn-binding site

LA site:

Low-affinity electron donation site

MBTH:

3-Methyl-2-benzothiazolinone hydrazone hydrochloride hydrate

OEC:

Oxygen-evolving complex

POBN:

4-Pyridyl-1-oxide-N-tert-butylnitrone

PSII:

Photosystem II

PSII(–Ca):

Ca2+-depleted PSII

PSII(–Ca, –Cl):

Ca2+- and Cl−-depleted PSII

PSII(–Mn):

Mn-depleted PSII

PSII(–PsbO):

PsbO-depleted PSII

RC:

Reaction center

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

YZ :

Redox-active tyrosine (D1-Tyr161), the first electron donor to P680 + in PSII

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Acknowledgments

The work at the National Renewable Energy Laboratory was carried out under US Department of Energy contract number DE-AC36-08-GO28308. This study was also supported in part by the Russian Foundation for Basic Research, Project Numbers 08-04-00354 (AR), by the Chemical Sciences, Geosciences and Biosciences Division, Office of Basic Energy Sciences, Office of Science, U.S. Department of Energy (MS), and by the NREL pension program (MS). The authors would like to thank Drs. Gary Brudvig and Ken Sauer for their valuable comments and recommendations during the inception of this work.

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Authors and Affiliations

  1. Department of Biophysics, Faculty of Biology, Lomonosov Moscow State University, 119234, Moscow, Russia

    Boris K. Semin, Lira N. Davletshina, Kirill N. Timofeev, Il’ya I. Ivanov & Andrei B. Rubin

  2. National Renewable Energy Laboratory, Energy Sciences Directorate, Golden, CO, 80401, USA

    Boris K. Semin & Michael Seibert

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  1. Boris K. Semin
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  2. Lira N. Davletshina
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  3. Kirill N. Timofeev
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Correspondence to Boris K. Semin.

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Semin, B.K., Davletshina, L.N., Timofeev, K.N. et al. Production of reactive oxygen species in decoupled, Ca2+-depleted PSII and their use in assigning a function to chloride on both sides of PSII. Photosynth Res 117, 385–399 (2013). https://doi.org/10.1007/s11120-013-9870-x

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