Two‐dimensional gel electrophoresis as tool for proteomics studies in combination with protein identification by mass spectrometry

Abstract

The proteome analysis by 2-DE is one of the most potent methods of analyzing the complete proteome of cells, cell lines, organs and tissues in proteomics studies. It allows a fast overview of changes in cell processes by analysis of the entire protein extracts in any biological and medical research projects. New instrumentation and advanced technologies provide proteomics studies in a wide variety of biological and biomedical questions. Proteomics work is being applied to study antibiotics-resistant strains and human tissues of various brain, lung, and heart diseases. It cumulated in the identification of antigens for the design of new vaccines. These advances in proteomics have been possible through the development of advanced high-resolution 2-DE systems allowing resolution of up to 10 000 protein spots of entire cell lysates in combination with protein identification by new highly sensitive mass spectrometric techniques. The present technological achievements are suited for a high throughput screening of different cell situations. Proteomics may be used to investigate the health effects of radiation and electromagnetic field to clarify possible dangerous alterations in human beings.

A proteomic approach to identifying proteins differentially expressed in conidia and mycelium of the entomopathogenic fungus Metarhizium acridum

“…Despite the limitations associated with current methodologies (Kniemeyer et al 2006;Wittmann-Liebold et al 2006), 2-DE proved to be a robust method for the study of changes in proteomes during fungal development. In the present study, without any enrichment strategies, approximately 1130 and 1200 protein spots were detected in Metarhizium acridum ungerminated conidia and fast-growing mycelia, respectively.…”

“…Proteomics allows qualitative and quantitative measurements of large numbers of proteins that directly influence cellular morphology and biochemistry, and thus provide accurate analysis of cellular protein status or changes during growth, differentiation and response to environmental factors (Wittmann-Liebold et al 2006;Bö hmer et al 2007;Kim et al 2007a, b;Kim et al 2008). Proteomics complements other functional genomics approaches such as transcriptomics, but it is a more reliable technique because proteins are directly related to the phenotype (Bhadauria et al 2007).…”

A CTAB based method for the preparation of total protein extract of wine spoilage microrganisms for proteomic analysis

“…Two-dimensional electrophoresis (2DE) is a powerful tool for the proteome analysis [1][2][3]. The technique, which separates proteins first on the basis of their charge (IEF) and then on their mass (SDS), has proven to be extremely effective to map most of the proteins expressed by a cell or a microorganism in a defined moment of its life.…”

Proteomics: A Successful Approach to Understand the Molecular Mechanism of Plant-Pathogen Interaction

“…After digestion with sequence specific proteases, the identity of proteins are analysed by mass spectrometry [17,18]. Analysis of 2D-PAGE gels simultaneously provides several pieces of informations such as molecular weight, isoelectric point, quantity as well as post translational modifications (PTMs) [15,19]. Various PTMs can be profiled via 2D-PAGE.…”