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Storage of light-driven transthylakoid proton motive force as an electric field (Δψ) under steady-state conditions in intact cells of Chlamydomonasreinhardtii

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Abstract

Proton motive force (pmf) is physiologically stored as either a ΔpH or a membrane potential (Δψ) across bacterial and mitochondrial energetic membranes. In the case of chloroplasts, previous work (Cruz et al. 2001, Biochemistry 40: 1226–1237) indicates that Δψ is a significant fraction of pmf, in vivo, and in vitro as long as the activities of counterions are relatively low. Kinetic analysis of light-induced changes in the electrochromic shift (ECS) in intact leaves was consistent with these observations. In this work, we took advantage of the spectroscopic properties of the green alga, Chlamydomonas reinhardtii, to demonstrate that light-driven Δψ was stored in vivo over the hours time scale. Analysis of the light-induced ECS kinetics suggested that the steady-state Δψ in 400 μmol photons m−2 s−1 red light was between 20 and 90 mV and that this represented about 60% of the light-induced increase in pmf. By extrapolation, it was surmised that about half of total (basal and light-induced) pmf is held as Δψ. It is hypothesized that Δψ is stabilized either by maintaining low chloroplast ionic strength or by active membrane ion transporters. In addition to the strong implications for regulation of photosynthesis by the xanthophyll cycle, these results imply that pmf partitioning is important across a wide range of species.

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Abbreviations

β:

buffering capacity

Δψinv:

inverted electric field component of pmf due to rapid proton efflux upon rapid light to dark transition

Δψ:

the electrical component of the proton motive force

ΔpH:

the osmotic component of the proton motive force

CCCP:

carbonyl m-chlorophenyl-hydrazone

DOFS:

diffused optics flash spectrophotometer

ECS:

electrochromic shift of light harvesting pigments

ECSd:

basal level of ECS in the dark

ECSinv:

the change in ECS upon a rapid light-dark transition, that extends below the baseline taken in the dark

ECSss:

the extent of light-induced ECS signal in the steady-state

ECSt:

the total difference in ECS signals upon shuttering the actinic light (ECSss–ECSinv)

HS medium:

high salt growth medium; LED, light emitting diode

P700+:

oxidized state of the chlorophyll special pair of photosystem I

pmf a :

threshold level of pmf required for activation of the ATP synthase

TAP medium:

tris-acetate-phosphate growth med- ium; pmf proton motive force

VDE:

violaxanthin de-epoxidase.

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  1. Institute of Biological Chemistry, Washington State University , Pullman, WA, 99164–6340, USA

    Jeffrey A. Cruz, Atsuko Kanazawa, Nathan Treff & David M. Kramer

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  1. Jeffrey A. Cruz
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Cruz, J.A., Kanazawa, A., Treff, N. et al. Storage of light-driven transthylakoid proton motive force as an electric field (Δψ) under steady-state conditions in intact cells of Chlamydomonasreinhardtii. Photosynth Res 85, 221–233 (2005). https://doi.org/10.1007/s11120-005-4731-x

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