Abstract
Inhibitory analysis is a useful tool for studying cytochrome b6f complex in the photosynthetic electron transport chain. Here, we examine the inhibitory efficiency of two widely used inhibitors of the plastoquinol oxidation in the cytochrome b6f complex, namely 2,4-dinitrophenyl ether of 2-iodo-4-nitrothymol (DNP-INT) and 2,5-dibromo-3-methyl-6-isopropylbenzoquinone (DBMIB). Using isolated thylakoids from pea and arabidopsis, we demonstrate that inhibitory activity of DNP-INT and DBMIB is enhanced by increasing irradiance, and this effect is due to the increase in the rate of electron transport. However, the accumulation of protons in the thylakoid lumen at low light intensity has opposite effects on the inhibitory activity of DNP-INT and DBMIB, namely increasing the activity of DNP-INT and restricting the activity of DBMIB. These results allow for the refinement of the conditions under which the use of these inhibitors leads to the complete inhibition of plastoquinol oxidation in the cytochrome b6f complex, thereby broadening our understanding of the operation of the cytochrome b6f complex under conditions of steady-state electron transport.
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Abbreviations
- Chl:
-
Chlorophyll a
- Cyt:
-
Cytochrome
- Cyt b 6 f :
-
Cytochrome b6f complex
- DBMIB:
-
2,5-Dibromo-3-methyl-6-isopropylbenzoquinone
- DCBQ:
-
2,6-Dichloro-1,4-benzoquinone
- DCMU:
-
3-(3,4-Dichlorophenyl)-1,1-dimethylurea
- DNP-INT:
-
2,4-Dinitrophenyl ether of 2-iodo-4-nitrothymol
- ETC:
-
Electron transport chain
- ETR:
-
Electron transport rate
- ETR0 :
-
Electron transport rate in the absence of inhibitors
- F :
-
Fluorescence
- GrD:
-
Gramicidin D
- HL:
-
High light
- IC 50 :
-
Half maximal inhibitory concentration
- ISP:
-
Iron-sulfur protein
- LL:
-
Low light
- MV:
-
Methyl viologen
- PSI, PSII:
-
Photosystem I, photosystem II respectively
- PQ, PQ• −, PQH2 :
-
Plastoquinone, plastosemiquinone, plastoquinol respectively
- Qo-site:
-
Quinol-oxidizing site at the Cyt b6f complex
- Qr-site:
-
Quinone-reducing site at the Cyt b6f complex
- SuIV:
-
Subunit IV
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Acknowledgements
The work was supported by Russian Scientific Foundation, project # 22-24-01074, https://rscf.ru/project/22-24-01074. The authors are thankful to Anna Krupyanko for the help with thylakoid membranes isolation and Dr. Maria Borisova-Mubarakshina (Institute of Basic Biological Problems) for the fruitful discussion. The authors are grateful to Professor Toshiharu Shikanai (Kyoto University) for kindly providing seeds of Col-5 and the pgr1 mutant.
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MK and BI: designed experiments; DV, IP, IN, and MK: performed experiments; MK: wrote the article with contributions from all the authors.
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Vilyanen, D., Pavlov, I., Naydov, I. et al. Peculiarities of DNP-INT and DBMIB as inhibitors of the photosynthetic electron transport. Photosynth Res (2023). https://doi.org/10.1007/s11120-023-01063-5
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DOI: https://doi.org/10.1007/s11120-023-01063-5