Journal
PLANT AND CELL PHYSIOLOGY
Volume 59, Issue 7, Pages 1353-1362Publisher
OXFORD UNIV PRESS
DOI: 10.1093/pcp/pcy069
Keywords
Arabidopsis; CE-MS; FT-ICR-MS; Metabolome; Proteome; Vacuole
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Funding
- Japan Science and Technology Corporation (JST) by CREST
- Ministry of Education, Culture, Sports, Science and Technology (MEXT) [22120006]
- Japan Society for Promotion of Science (JSPS) [16H04807]
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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.
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