Combinatorial peptide ligand libraries and plant proteomics: A winning strategy at a price |
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Authors: | Egisto Boschetti Laurence V Bindschedler Chaorong Tang Elisa Fasoli Pier Giorgio Righetti |
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Institution: | 1. Bio-Rad Laboratories, C/o CEA-Saclay-DSV iBiTec-S, 91181 Gif-sur-Yvette, France;2. The BioCentre and School of Biological, University of Reading, Whiteknights, Reading RG6 6AS, UK;3. Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Hainan 571737, China;4. Department of Chemistry, Materials and Chemical Engineering “Giulio Natta”, Politecnico di Milano, Via Mancinelli 7, Milan 20131, Italy |
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Abstract: | 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 / 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. |
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Keywords: | Plant proteomics Ligand library Hexapeptide ligands SELDI mass spectrometry Low-abundance proteome |
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