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Quantitative local photosynthetic flux measurements at isolated chloroplasts and thylakoid membranes using scanning electrochemical microscopy (SECM)
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McKelvey, Kim M. (Kim Martin), Martin, Sophie G., Robinson, Colin and Unwin, Patrick R. (2013) Quantitative local photosynthetic flux measurements at isolated chloroplasts and thylakoid membranes using scanning electrochemical microscopy (SECM). The Journal of Physical Chemistry Part B: Condensed Matter, Materials, Surfaces, Interfaces & Biophysical, Volume 117 (Number 26). pp. 7878-7888. doi:10.1021/jp403048f ISSN 1520-6106.
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WRAP_Urwin_1050272-ch-141013-isolated_chlorothy.pdf - Accepted Version Download (1575Kb) | Preview |
Official URL: http://dx.doi.org/10.1021/jp403048f
Abstract
Scanning electrochemical microscopy (SECM) offers a fast and quantitative method to measure local fluxes within photosynthesis. In particular, we have measured the flux of oxygen and ferrocyanide (Fe(CN)64–), from the artificial electron acceptor ferricyanide (Fe(CN)63–), using a stationary ultramicroelectrode at chloroplasts and thylakoid membranes (sourced from chloroplasts). Oxygen generation at films of chloroplasts and thylakoid membranes was detected directly during photosynthesis, but in the case of thylakoid membranes, this switched to sustained oxygen consumption at longer illumination times. An initial oxygen concentration spike was detected over both chloroplast and thylakoid membrane films, and the kinetics of the oxygen generation were extracted by fitting the experimental data to a finite element method (FEM) simulation. In contrast to previous work, the oxygen generation spike was attributed to the limited size of the plastoquinone pool, a key component in the linear electron transport pathway and a contributing factor in photoinhibition. Finally, the mobile nature of the SECM probe, and its high spatial resolution, also allowed us to detect ferrocyanide produced from a single thylakoid membrane. These results further demonstrate the power of SECM for localized flux measurements in biological processes, in this case photosynthesis, and that the high time resolution, combined with FEM simulations, allows the elucidation of quantitative kinetic information.
Item Type: | Journal Article | ||||
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Subjects: | Q Science > QC Physics Q Science > QD Chemistry |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) |
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Library of Congress Subject Headings (LCSH): | Chloroplasts, Thylakoids, Photosynthesis, Bioelectrochemistry, Electrochemistry, Microscopy | ||||
Journal or Publication Title: | The Journal of Physical Chemistry Part B: Condensed Matter, Materials, Surfaces, Interfaces & Biophysical | ||||
Publisher: | American Chemical Society | ||||
ISSN: | 1520-6106 | ||||
Official Date: | 2013 | ||||
Dates: |
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Volume: | Volume 117 | ||||
Number: | Number 26 | ||||
Page Range: | pp. 7878-7888 | ||||
DOI: | 10.1021/jp403048f | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published | ||||
Access rights to Published version: | Restricted or Subscription Access | ||||
Date of first compliant deposit: | 25 December 2015 | ||||
Date of first compliant Open Access: | 25 December 2015 | ||||
Funder: | Engineering and Physical Sciences Research Council (EPSRC), European Research Council (ERC), Advantage West Midlands (AWM), European Regional Development Fund (ERDF) | ||||
Grant number: | ERC-2009-AdG247143 QUANTIF (ERC) |
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