Sample preparation and fractionation for 2-D PAGE/Proteomics

摘要: The quality of a proteomic analysis of a cell compartment strongly depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific drawbacks:(i) the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP) during the isolation procedure;(ii) polysaccharide networks of cellulose, hemicelluloses and pectins form potential traps for contaminants such as intracellular proteins;(iii) the presence of proteins interacting in many different ways with the polysaccharide matrix require different procedures to elute them from the cell wall. Three categories of CWP are distinguished: labile proteins that have little or no interactions with cell wall components, weakly bound proteins extractable with salts, and strongly bound proteins. Two alternative protocols are decribed for cell wall proteomics:(i) non-destructive techniques allowing the extraction of labile …