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Proteomics analysis of plant tissues based on two-dimensional gel electrophoresis

Abstract:

Proteomics, either considered as being a scientific discipline or a methodological approach, will be presented from a historical perspective, as the heir of the classical protein chemistry or biochemistry. In the last 20 years, classical techniques for protein extraction and resolution such as electrophoresis and liquid chromatography have been continuously improved and implemented with those of mass spectrometry, gene sequencing, and bioinformatics tools, providing a new horizon in molecular biology research, and revealing the never-before imagined complexity of the protein world. The application of proteomics to plant biology research will be briefly discussed, focusing on descriptive and comparative studies and on two-dimensional electrophoresis-based platforms. There will be sections devoted to the different steps in a standard proteomics workflow: protein extraction, fractionation, electrophoresis, mass spectrometry, and data analysis. As each technique and protocol has to be adapted and optimized to the experimental system under investigation, rather than providing specific detailed protocols, general guidelines are presented, with reference to previous original publications and reviews from the author’s group and the current relevant literature. More than to scientists with real expertise in proteomics, this chapter is dedicated to plant physiologists, who aim to enter the field.