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The vacuolar membrane; a proteomics approach

Reference	BB/C510032/1
Principal Investigator / Supervisor	Professor Frans Maathuis
Co-Investigators / Co-Supervisors	Professor Dale Sanders
Institution	University of York
Department	Biology
Funding type	Research
Value (£)	262,700
Status	Completed
Type	Research Grant
Start date	01/02/2005
End date	30/06/2009
Duration	53 months

Abstract

Plant vacuoles play many crucial roles in plant growth and development. Proteins expressed in the vacuolar membrane, the tonoplast, are essential for the many storage and signalling functions of the vacuole and impact not only on plant growth but also on the nutritional value of plants as food. Many of the membrane transporters expressed in the tonoplast have not been identified at the gene level which greatly frustrates a proper functional characterisation. We propose to employ a proteomics approach to directly assess the toplast protein complement, through a membrane protein-optimised liquid chromatography, MALDI-TOF and ES-TOF tandem MS-MS methodology. This strategy will be used to separate and identify tonoplast membrane proteins, to carry out bioinformatics analyses on the tonoplast proteome and to identify putative proteins involved in Ca nutrition and Ca signalling. Isotope coded affinity tag (ICAT) labelling will be carried out on tonoplast fractions isolated in different Ca regimes to assess changes in protein expression and to integrate these proteomics data with previously obtained transcriptomics data from identical stress conditions. Further characterisation of important Ca signalling elements will be carried out using expression studies, post translational modification studies and phenotypic characterisation of null mutants.

Summary

unavailable

Committee	Closed Committee - Plant & Microbial Sciences (PMS)
Research Topics	Plant Science
Research Priority	X – Research Priority information not available
Research Initiative	Proteomics and Cell Function (PCF) [2003-2004]
Funding Scheme	X – not Funded via a specific Funding Scheme

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