Resolving subcellular plant metabolism

Author(s)
Lisa Fuertauer, Lisa Kuestner, Wolfram Weckwerth, Arnd G. Heyer, Thomas Naegele
Abstract

Plant cells are characterized by a high degree of compartmentalization and a diverse proteome and metabolome. Only a very limited number of studies has addressed combined subcellular proteomics and metabolomics which strongly limits biochemical and physiological interpretation of large-scale 'omics data. Our study presents a methodological combination of nonaqueous fractionation, shotgun proteomics, enzyme activities and metabolomics to reveal subcellular diurnal dynamics of plant metabolism. Subcellular marker protein sets were identified and enzymatically validated to resolve metabolism in a four-compartment model comprising chloroplasts, cytosol, vacuole and mitochondria. These marker sets are now available for future studies that aim to monitor subcellular metabolome and proteome dynamics. Comparing subcellular dynamics in wild type plants and HXK1-deficient gin2-1 mutants revealed a strong impact of HXK1 activity on metabolome dynamics in multiple compartments. Glucose accumulation in the cytosol of gin2-1 was accompanied by diminished vacuolar glucose levels. Subcellular dynamics of pyruvate, succinate and fumarate amounts were significantly affected in gin2-1 and coincided with differential mitochondrial proteome dynamics. Lowered mitochondrial glycine and serine amounts in gin2-1 together with reduced abundance of photorespiratory proteins indicated an effect of the gin2-1 mutation on photorespiratory capacity. Our findings highlight the necessity to resolve plant metabolism to a subcellular level to provide a causal relationship between metabolites, proteins and metabolic pathway regulation.

Organisation(s)
Department of Ecogenomics and Systems Biology, Functional and Evolutionary Ecology, Large-Instrument Facility for Mass Spectrometry in Life Sciences, Research Platform Vienna Metabolomics Center
External organisation(s)
Ludwig-Maximilians-Universität München, Universität Stuttgart
Journal
The Plant Journal
Volume
100
Pages
438-455
No. of pages
18
ISSN
0960-7412
DOI
https://doi.org/10.1111/tpj.14472
Publication date
10-2019
Peer reviewed
Yes
Austrian Fields of Science 2012
Cell biology, Metabolomics , Biochemistry
Keywords
ASJC Scopus subject areas
Portal url
https://ucris.univie.ac.at/portal/en/publications/resolving-subcellular-plant-metabolism(dd3f7140-4216-4b0b-b68f-1901dd0df91a).html