Two-dimensional gel electrophoresis in bacterial proteomics

查看全文 作　　者：Shirly [1]O.T.Curreem;Rory [2]M.Watt;Susanna [1,3,4,5]K.P.Lau;Patrick [1,3,4,5]C.Y.Woo

机构地区：[1]Department of Microbiology,The University of Hong Kong,Queen Mary Hospital,Hong Kong,China;[2]Oral Biosciences,Faculty of Dentistry,The University of Hong Kong,Queen Mary Hospital,Hong Kong,China;[3]State Key Laboratory of Emerging Infectious Diseases,Department of Microbiology,The University of Hong Kong,Queen Mary Hospital,Hong Kong,China;[4]Research Centre of Infection and Immunology,The University of Hong Kong,Queen Mary Hospital,Hong Kong,China;[5]Carol Yu Centre of Infection,The University of Hong Kong,Hong Kong,China

出　　处：《Protein & Cell》索引2012年第3卷第5期,共18页

基　　金：This work was partly supported by the Research Fund for the Control of Infectious Diseases(Commissioned study)of the Health,Welfare and Food Bureau of the Hong Kong SAR Government,Research Grant Council Grant;University Development Fund. 摘　　要：Two-dimensional gel electrophoresis(2-DE)is a gel-based technique widely used for analyzing the pro-tein composition of biological samples.It is capable of resolving complex mixtures containing more than a thousand protein components into individual protein spots through the coupling of two orthogonal bio-physical separation techniques:isoelectric focusing(first dimension)and polyacrylamide gel electrophore-sis(second dimension).2-DE is ideally suited for ana-lyzing the entire expressed protein complement of a bacterial cell:its proteome.Its relative simplicity and good reproducibility have led to 2-DE being widely used for exploring proteomics within a wide range of envi-ronmental and medically-relevant bacteria.Here we give a broad overview of the basic principles and his-torical development of gel-based proteomics,and how this powerful approach can be applied for studying bacterial biology and physiology.We highlight specific 2-DE applications that can be used to analyze when,where and how much proteins are expressed.The links between proteomics,genomics and mass spectrometry are discussed.We explore how proteomics involving tandem mass spectrometry can be used to analyze(post-translational)protein modifications or to identify proteins of unknown origin by de novo peptide se-quencing.The use of proteome fractionation tech-niques and non-gel-based proteomic approaches are also discussed.We highlight how the analysis of pro-teins secreted by bacterial cells(secretomes or exo-proteomes)can be used to study infection processes or the immune response.This review is aimed at non-specialists who wish to gain a concise,compre-hensive and contemporary overview of the nature and applications of bacterial proteomics. 关键词：two-dimensional gel electrophoresis bacte-ria PROTEOMICS

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Two-dimensional gel electrophoresis in bacterial proteomics

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