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Combinatorial peptide ligand libraries and plant proteomics: A winning strategy at a price

文献类型： 外文期刊

作者： Boschetti, Egisto 2 ; Bindschedler, Laurence V. 3 ; Tang, Chaorong 4 ; Fasoli, Elisa 1 ; Righetti, Pier Giorgio 1 ;

作者机构： 1.Politecn Milan, Dept Chem Mat & Chem Engn Giulio Natta, I-20131 Milan, Italy

2.CEA Saclay DSV IBITec S, Biorad Labs, F-91181 Gif Sur Yvette, France

3.Univ Reading, BioCtr, Reading RG6 6AS, Berks, England

4.Chinese Acad Trop Agr Sci, Rubber Res Inst, Hainan 571737, Peoples R China

关键词： Plant proteomics;Ligand library;Hexapeptide ligands;SELDI mass spectrometry;Low-abundance proteome;MASS-SPECTROMETRY;ARABIDOPSIS-THALIANA;GEL-ELECTROPHORESIS;PROTEINS;SYSTEM;PERSPECTIVES;TECHNOLOGY;TRENDS;BEADS;IMPURITIES

期刊名称：JOURNAL OF CHROMATOGRAPHY A （ 影响因子：4.759； 五年影响因子：4.302 ）

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收录情况： SCI

摘要： The mechanism of action and properties of a solid-phase ligand library made of hexapeptides (combinatorial peptide ligand libraries or CPLL, for capturing the "hidden proteome", i.e. the low- and very low-abundance proteins Constituting the vast majority of species in any proteome. as applied to plant tissues, are reviewed here. Plant tissues are notoriously recalcitrant to protein extraction and to proteome analysis, Firstly, rigid plant cell walls need to be mechanically disrupted to release the cell content and, in addition to their poor protein yield, plant tissues are rich in proteases and oxidative enzymes, contain phenolic Compounds, starches, oils, pigments and secondary metabolites that massively contaminate protein extracts. In addition, complex matrices of polysaccharides, including large amount of anionic pectins, are present. All these species compete with the binding of proteins to the CPLL beads, impeding proper capture and identification I detection of low-abundance species. When properly pre-treated, plant tissue extracts are amenable to capture by the CPLL beads revealing thus many new species among them low-abundance proteins. Examples are given on the treatment of leaf proteins, of corn seed extracts and of exudate proteins (latex from Hevea brasiliensis). In all cases, the detection of unique gene products via CPLL Capture is at least twice that of control, untreated sample. (c) 2008 Elsevier B.V. All rights reserved.

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