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Plant Cell Physiol. 2018 Jul 1;59(7):1353-1362. doi: 10.1093/pcp/pcy069.

Molecular Components of Arabidopsis Intact Vacuoles Clarified with Metabolomic and Proteomic Analyses.

Author information

1
Department of Biology, Graduate School of Science, Kobe University, Nada, Kobe, Japan.
2
CREST, JST, Sanbancho Bldg, 5 Sanban-cho, Chiyoda-ku, Tokyo, Japan.
3
Department of Science, Okayama University of Science, Okayama, Japan.
4
Graduate School of Pharmaceutical Science, Osaka University, Suita, Japan.
5
Faculty of Agriculture, Yamagata University, Tsuruoka, Japan.
6
RIKEN Center for Sustainable Resource Science, Yokohama, Japan.
7
Graduate School of Engineering, Osaka University, Suita, Japan.
8
Graduate School of Information Science and Technology, Osaka University, Suita, Japan.
9
Graduate School of Information Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara, Japan.
10
Department of Chemistry and Biochemistry, Old Dominion University, Norfolk, VA, USA.
11
Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Naka-ku, Sakai, Japan.
12
Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan.

Abstract

We analyzed the metabolites and proteins contained in pure intact vacuoles isolated from Arabidopsis suspension-cultured cells using capillary electrophoresis-mass spectrometry (CE-MS), Fourier transform-ion cyclotron resonance (FT-ICR)-MS and liquid chromatography (LC)-MS. We identified 21 amino acids and five organic acids as major primary metabolites in the vacuoles with CE-MS. Further, we identified small amounts of 27 substances including well-known vacuolar molecules, but also some unexpected substances (e.g. organic phosphate compounds). Non-target analysis of the vacuolar sample with FT-ICR-MS suggested that there are 1,106 m/z peaks that could predict the 5,090 molecular formulae, and we have annotated 34 compounds in these peaks using the KNapSAck database. By conducting proteomic analysis of vacuolar sap, we found 186 proteins in the same vacuole samples. Since the vacuole is known as a major degradative compartment, many of these were hydrolases, but we also found various oxidoreductases and transferases. The relationships between the proteins and metabolites in the vacuole are discussed.

PMID:
29660082
DOI:
10.1093/pcp/pcy069
[Indexed for MEDLINE]

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