The proteomics support facility will provide technological innovations, training of expert staff in biological mass spectrometry and protein micro-characterization service. The facility is equipped with all the tools required for proteomics studies: state of the art mass spectrometers, N-terminal sequencer, 2D gel electrophoresis facility and bioinformatics. Mass spectrometry will be the main tool for the Consortium studies because it can provide a wide variety of information about protein structure requiring only small amounts of protein for analysis, thus enabling effective interface with molecular biology techniques. The goal of the Protein Microsequencing Facility is to assist scientists in the NoE with their research by helping them to characterize proteins of interest. In selected cases the facility provides access to equipment and technical expertise on a basis which is much more cost effective than establishing the techniques in each of the user's laboratories.
The Proteomics Facility is involved in R&D projects focusing on the development and application of sensitive and specific methods for protein identification and microcharacterization. In particular, they are developing novel approaches that can be applied to proteome analysis of cells under physiological and pathological states. They are using biological mass spectrometry for the identification of proteins and multiprotein complexes aimed at revealing protein-protein interactions; and for the identification of their post-translational modifications.
Mass spectrometry is the principal analytical tool used by the Facility, as this technique is, up to now, the most sensitive and specific tool for protein microanalysis. Methods for in solution and in gel digestion have been established and successful analysis of proteins and protein complexes separated on commassie or silver stained gels is now possible. The Facility can perform, on a routine basis, peptide mass mapping and tandem MS experiments by on line and off line nanoelectrospray. In the last three years the Facility has set up specific and sensitive methods for the enrichment and the isolation of phosphorylated peptides present in in gel separated proteins. Moreover, to assess the oxidative status of cysteine residues, they have recently developed a new method based on in gel digestion, on target reduction and alkylation of the cysteine residues and on MALDI-TOF MS, which allows the rapid and sensitive characterization of cysteine contating proteins.