Features of using 2,6-dichlorophenolindophenol as an electron acceptor in photosynthesis studies
Abstract
2,6-dichlorophenolindophenol (DCPIP) is a redox indicator widely used to study electron transfer reactions in biological systems, including in the process of photosynthesis. DCPIP exists in solution in two forms – “pink” and “blue,” which transform into each other during protonation/deprotonation. Upon reduction, the DCPIP is discolored. We investigated the pH-dependence of DCPIP reduction rate in the presence of the photosystem II (PSII) at two wavelengths – 522 nm (isobestic DCPIP point) and 600 nm (near the absorption maximum of the deprotonated “blue” form). It was shown that in experiments with change of the pH medium, measuring at a wavelength of 600 nm requires corrections related to changing the ratio of the “blue” and “pink” forms of the acceptor, as well as using the pK parameter of this acceptor, whose values рК vary in various sources, to calculate the DCPIP reduction rate. Measurements at the isobestic point (522 nm) avoid these complexities. We also found that the maximum at the pH-dependence of the DCPIP reduction rate by PSII shifted by about 1 unit to the acidic region relative to the maximum of the acceptor pair 2,6-dichloro-p-benzoquinone – potassium ferricyanide reduction rate pH-dependence. This shift may be due to the lower availability of the QB site on the acceptor side PSII for the charged deprotonated DCPIP form compared to the uncharged protonated form.
Keywords
2,
6-dichlorophenolindophenol,
photosystem II,
pH-dependence,
electron acceptors,
2,
6-dichloro-p-benzoquinone,
potassium ferricyanide,
photometry
Supporting agencies: The research was carried out as part of the Scientific Project of the State Order of the Government of Russian Federation to Lomonosov Moscow State University No. 121032500058-7
About the Authors
A. V. Loktyushkin
Lomonosov Moscow State University
Russian Federation
Russian Federation
Department of Biophysics, School of Biology
Leninskiye gory 1–12, Moscow 119234
E. R. Lovyagina
Lomonosov Moscow State University
Russian Federation
Russian Federation
Department of Biophysics, School of Biology
Leninskiye gory 1–12, Moscow 119234
B. K. Semin
Lomonosov Moscow State University
Russian Federation
Russian Federation
Department of Biophysics, School of Biology
Leninskiye gory 1–12, Moscow 119234
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Review
For citations:
Loktyushkin A.V., Lovyagina E.R., Semin B.K. Features of using 2,6-dichlorophenolindophenol as an electron acceptor in photosynthesis studies. Vestnik Moskovskogo universiteta. Seriya 16. Biologiya. 2021;76(4):234-240. (In Russ.)
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