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2-D HPLC coupled to ion trap mass spectrometry functional proteomics and the analysis of postranslational protein modifications

Reference	JE616379
Principal Investigator / Supervisor	Dr Josip Lovric
Co-Investigators / Co-Supervisors	Professor Mark Peter Ashe, Dr Graeme Conn, Professor Christopher Grant, Professor John Hyde, Professor John McCarthy, Professor Graham Pavitt, Dr Paul Sims
Institution	The University of Manchester
Department	Life Sciences
Funding type	Research
Value (£)	194,108
Status	Completed
Type	Research Grant
Start date	27/06/2002
End date	27/06/2003
Duration	12 months

Abstract

We apply for funds to purchase a nano HPLC system and an ion trap mass spectrometer for high sensitivity 2D- HPLC-MS to analyse proteins and peptides from complex mixtures for: Sequencing proteins of interest in order to determine or verify identity; Mapping of posttranslational modifications in proteins with high sensitivity in medium throughput; Characterisation of protein signalling complexes and protein/RNA interactions; Identification of differentially expressed/modified proteins in conjunction with 2-D gel and Free Flow electrophoresis based expression profiling in proteomics with high throughput; and Development of protocols for the replacement of gels in functional proteomics. The projects that will exploit this system mainly involve research in biomedical sciences of pathogenic species and mammalian cells, biotechnological applications and research on fundamental cellular regulation in model organisms such as yeast. The instrumentation to be purchased (LC Packings nano-HPLC/Thermo Finnigan ion trap) allows these projects to take advantage of proteomics approaches. It will also be used to develop alternative analysis methods to traditional 2-D gel electrophoresis for small, hydrophobic/membrane and large proteins. This will involve a combination of Free Flow Electrophoresis technology (funded by the DTI) with 2-D HPLC MS. The instrumentation is highly complementary to the facilities at the Michael Barber Centre for Mass Spectroscopy and the LRF proteomics facility at UMIST, and will be accessible to researchers for all Departments at UMIST.

Summary

unavailable

Committee	Closed Committee - Biochemistry & Cell Biology (BCB)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	Joint Equipment Initiative 2001 (JE6) [2001]
Funding Scheme	X – not Funded via a specific Funding Scheme

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