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High sensitivity - high mass spectrometry in the structural studies of biomolecules

Objective


The major research projects done by SESMA with the European Mass Spectrometry Network included:

- Analysis of human haemoglobin variants. This technique is used to give chemical and molecular descriptions of diseases. Using this procedure, a large number of haemoglobins were characterised, including several uncommon and two completely new genetic variants.

- Characterisation of protein adducts with genotoxic xenobiotics. To help detect cancer risk with better certainty, this research aimed at gathering more and better quality information about the reactivity of genotoxic xenobiotics with proteins and peptides, using mass spectrometry.

- Post-translational modifications. The accurate determination of protein molecular weight by electrospray mass spectrometry can provide definitive data on the occurrence of post-translational events, including proteolytic processing of the polypeptide chain. In this case, the accuracy of the analysis leads to the location of the maturation site within the protein sequence.

- Recombinant protein analysis. The application of recombinant DNA methodologies greatly enhanced the need for definitive structural information on protein products obtained by genetic engineering. The most important alternative approach to classic biochemistry procedures (Edman degradation, HPLC mapping, etc) is the use of mass spectrometry methods such as ES/MS, FAB/MS and FAB/MS/MS.
This project helped form a European Network on Biomolecular Mass Spectrometry involving eight laboratories from across the EU, working in the field of biological mass spectrometry.

Funding for this project coincided with the purchase, by the National Research Council, of a ZAB-T tandem mass spectrometer allowing high sensitivity, high mass four sectors (tandem), characterised by a new geometry and designed to address the structural analysis of biomolecules in the low femtomole range. This device combines the most recent innovations in the production and detection of high mass ions, which play an increasing role in the determination of peptide and other biopolymer molecular weights.

Coordinator

Consiglio Nazionale delle Ricerche (CNR)
Address
111,Via P. Castellino
80131 Napoli
Italy