Protein-protein interactions and metabolite channelling in the plant tricarboxylic acid cycle

Youjun Zhang, Katherine F. M. Beard, Corné Swart, Susan Bergmann, Ina Krahnert, Zoran Nikoloski, Alexander Graf, R. George Ratcliffe, Lee J. Sweetlove, Alisdair R. Fernie () and Toshihiro Obata
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Youjun Zhang: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Katherine F. M. Beard: University of Oxford
Corné Swart: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Susan Bergmann: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Ina Krahnert: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Zoran Nikoloski: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Alexander Graf: Max-Planck-Institut für Molekulare Pflanzenphysiologie
R. George Ratcliffe: University of Oxford
Lee J. Sweetlove: University of Oxford
Alisdair R. Fernie: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Toshihiro Obata: Max-Planck-Institut für Molekulare Pflanzenphysiologie

Nature Communications, 2017, vol. 8, issue 1, 1-11

Abstract: Abstract Protein complexes of sequential metabolic enzymes, often termed metabolons, may permit direct channelling of metabolites between the enzymes, providing increased control over metabolic pathway fluxes. Experimental evidence supporting their existence in vivo remains fragmentary. In the present study, we test binary interactions of the proteins constituting the plant tricarboxylic acid (TCA) cycle. We integrate (semi-)quantitative results from affinity purification-mass spectrometry, split-luciferase and yeast-two-hybrid assays to generate a single reliability score for assessing protein–protein interactions. By this approach, we identify 158 interactions including those between catalytic subunits of sequential enzymes and between subunits of enzymes mediating non-adjacent reactions. We reveal channelling of citrate and fumarate in isolated potato mitochondria by isotope dilution experiments. These results provide evidence for a functional TCA cycle metabolon in plants, which we discuss in the context of contemporary understanding of this pathway in other kingdoms.

Date: 2017
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DOI: 10.1038/ncomms15212

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