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LC-MS facilities for analysis of secondary metabolites peptides and protein complexes in plants pests and pathogens

Reference	REI18496
Principal Investigator / Supervisor	Professor John Mansfield
Co-Investigators / Co-Supervisors	Professor Murray Grant, Dr John Rossiter, Dr Po-Yuan Shih
Institution	Imperial College London
Department	Biological Sciences
Funding type	Research
Value (£)	199,692
Status	Completed
Type	Research Grant
Start date	01/11/2002
End date	01/10/2003
Duration	11 months

Abstract

We plan to develop a multi-user LC-MS facility at Imperial College, Wye, which will form an integral part of present and future BBSRC research programmes. Both metabolomics and proteomics projects will be supported by the Applied Biosystems QTRAP instrument which combines the robust ease of usage of ion trap spectrometers with the sensitivity and mass accuracy of triple quadruple technology. The instrument will be established with both LC and capillary LC interfaces and will support projects in four main areas, each of which currently receives BBSRC or other grant support: 1) Plant-microbe interactions - comparative proteomics of the Arabidopsis/Pseudomomas syringae interaction, resistance gene signalling networks, effector targeting to protein complexes, and structure of the type III secretion apparatus; 2) Long distance signalling - comparative metabolomics and proteomics of transport pathway constituents, identification of signals for branching; 3) Plant- insect interactions - glucosinolate modification in transgenic Arabidopsis, pest responses, and salicylate dependent feeding determinants; 4) Plant secondary metabolites - biosynthesis and metabolite profiling of sesquiterpenoids in lettuce, and flavonol profiles in processed onion and kale. For the proteomics applications peptide sequences determined by LC-MS/MS will be used to interrogate Arabidopsis and other public databases for protein identification and to determine post-translational modification. Quantitative comparative proteomics will be developed using isotope coded affinity tags (ICAT) as a method to determine relative amounts of protein components. In the analysis of plant hormones and secondary metabolites, in addition to qualitative LC-MS profiling, the sensitivity afforded by the QTRAP will allow quantitative analysis of low abundance metabolites via internal standard isotope ratio mass spectrometry.

Summary

unavailable

Committee	Closed Committee - Plant & Microbial Sciences (PMS)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	Research Equipment Initiative 2002 (REI) [2002]
Funding Scheme	X – not Funded via a specific Funding Scheme

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