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Reduction of photosystem I reaction center by recombinant DrgA protein in isolated thylakoid membranes of the cyanobacterium Synechocystis sp. PCC 6803

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Abstract

To study the function of soluble NAD(P)H:quinone oxidoreductase of the cyanobacterium Synechocystis sp. PCC 6803 encoded by drgA gene, recombinant DrgA protein carrying 12 histidine residues on the C-terminal end was expressed in Escherichia coli and purified. Recombinant DrgA is a flavoprotein that exhibits quinone reductase and nitroreductase activities with NAD(P)H as the electron donor. Using EPR spectroscopy, it was demonstrated that addition of recombinant DrgA protein and NADPH to DCMU-treated isolated thylakoid membranes of the cyanobacterium increased the dark rereduction rate of the photosystem I reaction center (P700+). Thus, DrgA can participate in electron transfer from NADPH to the electron transport chain of the Synechocystis sp. PCC 6803 thylakoid membrane.

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Abbreviations

DCMU:

3-(3,4-dichlorophenyl)-1,1-dimethyl-urea

DCPIP:

2,6-dichlorophenol indophenol; decylplasto-quinone, 2,3-dimethyl-6-decyl-1,4-benzoquinone; dinoseb, 2-sec-butyl-4,6-dinitrophenol; duroquinone, tetramethyl-1,4-benzoquinone

ETC:

electron transport chain

Fd:

ferredoxin

FMN:

flavin mononucleotide

FNR:

ferredoxin:NADP+-oxidoreductase

IPTG:

isopropyl-β-D-thiogalactopyranoside

PCR:

polymerase chain reaction

PMSF:

phenylmethylsulfonyl fluoride

PQ:

plastoquinone

PS:

photosystem

Q0 :

2,3-dimethoxy-5-methyl-1,4-benzoquinone (ubiquinone Q0)

SOD:

superoxide dismutase

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Authors and Affiliations

  1. Faculty of Biology, Lomonosov Moscow State University, 119992, Moscow, Russia

    I. V. Elanskaya, V. G. Grivennikova, E. M. Muronets, E. P. Lukashev & K. N. Timofeev

  2. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997, Moscow, Russia

    V. A. Toporova

Authors
  1. I. V. Elanskaya
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  2. V. A. Toporova
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  3. V. G. Grivennikova
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  4. E. M. Muronets
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  5. E. P. Lukashev
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  6. K. N. Timofeev
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Corresponding author

Correspondence to I. V. Elanskaya.

Additional information

Original Russian Text © I. V. Elanskaya, V. A. Toporova, V. G. Grivennikova, E. M. Muronets, E. P. Lukashev, K. N. Timofeev, 2009, published in Biokhimiya, 2009, Vol. 74, No. 10, pp. 1327–1335.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM08-368, June 14, 2009.

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Elanskaya, I.V., Toporova, V.A., Grivennikova, V.G. et al. Reduction of photosystem I reaction center by recombinant DrgA protein in isolated thylakoid membranes of the cyanobacterium Synechocystis sp. PCC 6803. Biochemistry Moscow 74, 1080–1087 (2009). https://doi.org/10.1134/S0006297909100034

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  • DOI: https://doi.org/10.1134/S0006297909100034

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