Selected Bioinformatic Tools and MS (MALDI-TOF, PMF) Techniques Used in the Strategy for the Identification of Oat...

Computer analysis of protein maps obtained from the separation of proteins with two-dimensional polyacrylamide gel electrophoresis (2-DE), in combination with mass spectrometry (MS) analysis and selected bioinformatic tools is used in the strategy for the identification of oat proteins. In proteomic research the most often used MS technique is the combination of ion sources: matrix-assisted laser desorption/ionization (MALDI) and the analyzer of the time of flight (TOF), i.e., MALDI-TOF MS.This chapter describes the possibilities of the use of selected bioinformatic tools (UniProtKB database, ProtParam, Compute pI/MW programs) for initial identification of separated oat proteins (especially prolamin fractions) with the 2-DE technique. Also the procedure of preparation of samples obtained from cut out protein spots for analysis with the MALDI-TOF MS and peptide mass fingerprinting (PMF) technique is presented.Among oat prolamins separated with the 2-DE technique (see Chapter 17 ), 13 protein spots are considered to be the most characteristic (range of MW 27.0-34.6 kDa, pI 5.7-7.6) for this fraction of proteins. Among them there are four protein spots (MW 27.0-28.0 kDa) and two spots (MW 31.4-32.1 kDa) which can correspond to avenins (Accession numbers (AC) in UniProtKB: L0L5I0, I4EP88, I4EP64, L0L4I8 and F2Q9W5, L0L6J0, respectively).