Protein Characterization by Proteomics and Mass Spectrometry-Based Approaches
In the last decades, the rapid technological development in molecular biology together with the advances in large-scale protein analysis has contributed to move the attention of scientists from the characterization of individual proteins to a detailed examination of the entire proteome. New protein analytical strategies have emerged and mass spectrometry has become an indispensable tool in proteomic studies. From an analytical point of view, proteomic investigations are based on the use of highly efficient separation techniques for proteins such as reversed phase high performance liquid chromatography (RP-HPLC) or two dimensional polyacrylamide gel electrophoresis (2D-PAGE) combined with biochemical methods, enzymatic digestion and mass spectrometric methods in order to obtain data suitable for searching, by specific software (e.g. Mascot, PEAKS etc.) protein databases. As general statement, the overall goal of proteomics is to understand the function of all proteins present in an organism. This aim implies proteins identification together with the systematic determination of their different properties, including quantification, characterization of primary structure and post-translational modifications (PTMs), determination of protein interactions and sub-cellular distribution. Within this frame, the Laboratory of Organic Mass Spectrometry (Laboratorio di Spettrometria di Massa Organica, LSMO) is currently active in the development of MS-based strategies for the characterizations of the structure and post-translational modifications of the mitochondrial membrane proteins, milk proteins and proteins from wheat kernels. In detail, studies will continue on the characterization by mass spectrometry of the structure and post-translational modifications of the mitochondrial membrane proteins VDACs (voltage-dependent anion channels), in order to contribute to the definition of the function of these proteins, which play a determining role in the regulation of the exchange of metabolites between the mitochondria and the cell. The activity of the Lab will also include the characterization of milk proteins. Particularly, proteomic investigation of major and minor components already carried out for equine, ovine and caprine species will be extended to the milk of other mammals. This investigation, as well as the identification of proteins with allergenic characteristics, will be aimed at the identification of bioactive proteins and peptides with nutraceutical properties. Additionally, the research activity will be focused on the analysis at both qualitative and quantitative level of the protein fractions extracted from mature kernel of ancient and modern (improved varieties) durum wheats, with particular attention to components with allergenic characteristics (anti-nutritional factors). The results of these investigations will contribute to understand the relationship occurring between protein profile of ancient wheat varieties and potential benefits for human consumption.