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dc.contributor.authorArrivault, Stéphanie
dc.contributor.authorGuenther, Manuela
dc.contributor.authorFlorian, Alexandra
dc.contributor.authorEncke, Beatrice
dc.contributor.authorFeil, Regina
dc.contributor.authorVosloh, Daniel
dc.contributor.authorLunn, John E.
dc.contributor.authorSulpice, Ronan
dc.contributor.authorFernie, Alisdair R.
dc.contributor.authorStitt, Mark
dc.contributor.authorSchulze, Waltraud X.
dc.date.accessioned2018-09-20T16:00:09Z
dc.date.available2018-09-20T16:00:09Z
dc.date.issued2014-05-27
dc.identifier.citationArrivault, Stéphanie; Guenther, Manuela; Florian, Alexandra; Encke, Beatrice; Feil, Regina; Vosloh, Daniel; Lunn, John E. Sulpice, Ronan; Fernie, Alisdair R.; Stitt, Mark; Schulze, Waltraud X. (2014). Dissecting the subcellular compartmentation of proteins and metabolites in arabidopsis leaves using non-aqueous fractionation. Molecular & Cellular Proteomics 13 (9), 2246-2259
dc.identifier.issn1535-9476,1535-9484
dc.identifier.urihttp://hdl.handle.net/10379/10294
dc.description.abstractNon-aqueous fractionation is a technique for the enrichment of different subcellular compartments derived from lyophilized material. It was developed to study the subcellular distribution of metabolites. Here we analyzed the distribution of about 1,000 proteins and 70 metabolites, including 22 phosphorylated intermediates in wild-type Arabidopsis rosette leaves, using non-aqueous gradients divided into 12 fractions. Good separation of plastidial, cytosolic, and vacuolar metabolites and proteins was achieved, but cytosolic, mitochondrial, and peroxisomal proteins clustered together. There was considerable heterogeneity in the fractional distribution of transcription factors, ribosomal proteins, and subunits of the vacuolar-ATPase, indicating diverse compartmental location. Within the plastid, sub-organellar separation of thylakoids and stromal proteins was observed. Metabolites from the Calvin-Benson cycle, photorespiration, starch and sucrose synthesis, glycolysis, and the tricarboxylic acid cycle grouped with their associated proteins of the respective compartment. Nonaqueous fractionation thus proved to be a powerful method for the study of the organellar, and in some cases sub-organellar, distribution of proteins and their association with metabolites. It remains the technique of choice for the assignment of subcellular location to metabolites in intact plant tissues, and thus the technique of choice for doing combined metabolite-protein analysis on a single tissue sample.
dc.publisherAmerican Society for Biochemistry & Molecular Biology (ASBMB)
dc.relation.ispartofMolecular & Cellular Proteomics
dc.subjectcastor bean endosperm
dc.subjecttandem mass-spectrometry
dc.subjectenzyme-activities
dc.subjectbarley leaves
dc.subjectamino-acids
dc.subject2-dimensional electrophoresis
dc.subjectliquid-chromatography
dc.subjectstarch synthesis
dc.subjectspinach leaves
dc.subjectproteomics
dc.titleDissecting the subcellular compartmentation of proteins and metabolites in arabidopsis leaves using non-aqueous fractionation
dc.typeArticle
dc.identifier.doi10.1074/mcp.m114.038190
dc.local.publishedsourcehttp://www.mcponline.org/content/13/9/2246.full.pdf
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