Interactions of photosystem II with bicarbonate, formate and acetate

• In this study, we probe the effects of bicarbonate (hydrogencarbonate), BC, removal from photosystem II in spinach thylakoids by measuring flash-induced oxygen evolution patterns (FIOPs) with a Joliot-type electrode. For this we compared three commonly employed methods: (1) washing in BC-free medium, (2) formate addition, and (3) acetate addition. Washing of the samples with buffers depleted of BC and CO2 by bubbling with argon (Method 1) under our conditions leads to an increase in the double hit parameter of the first flash (beta(1)), while the miss parameter and the overall activity remain unchanged. In contrast, addition of 40-50 mM formate or acetate results in a significant increase in the miss parameter and to an similar to 50% (formate) and similar to 10% (acetate) inhibition of the overall oxygen evolution activity, but not to an increased beta(1) parameter. All described effects could be reversed by washing with formate/acetate free buffer and/or addition of 2-10 mM bicarbonate. The redox potential of the water-oxidizing complex (WOC) in samples treated by Method 1 is compared to samples containing 2 mM bicarbonate in two ways: (1) The lifetimes of the S-0, S-2, and S-3 states were measured, and no differences were found between the two sample types. (2) The S-1, S-0, S-1, and S-2 states were probed by incubation with small concentrations of NH2OH. These experiments displayed a subtle, yet highly reproducible difference in the apparent S-i/S-i state distribution which is shown to arise from the interaction of BC with PSII in the already reduced states of the WOC. These data are discussed in detail by also taking into account the CO2 concentrations present in the buffers after argon bubbling and during the measurements. These values were measured by membrane-inlet mass spectrometry (MIMS).

Ämnesord

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Nyckelord

flash-induced oxygen evolution pattern (FIOPs)

membrane-inlet mass spectrometry (MIMS)

S states

water splitting

oxygen evolution

bicarbonate

hydrogencarbonate

acetate

formate

Publikations- och innehållstyp

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