Farinazzo Alessia, Restuccia Umberto, Bachi Angela, Guerrier Luc, Fortis Frederic, Boschetti Egisto, Fasoli Elisa, Citterio Attilio, Righetti Pier Giorgio
Department of Chemistry, Materials and Chemical Egineering Giulio Natta, Politecnico di Milano, Via Mancinelli 7, 20131 Milan, Italy.
J Chromatogr A. 2009 Feb 20;1216(8):1241-52. doi: 10.1016/j.chroma.2008.11.051. Epub 2008 Nov 27.
The use of combinatorial peptide ligand libraries (CPLLs), containing hexapeptides terminating with a primary amine, or modified with a terminal carboxyl group, or with a terminal tertiary amine, allowed discovering and identifying a large number of previously unreported egg yolk proteins. Whereas the most comprehensive list up to date [K. Mann, M. Mann, Proteomics, 8 (2008) 178-191] tabulated about 115 unique gene products in the yolk plasma, our findings have more than doubled this value to 255 unique protein species. From the initial non-treated egg yolk it was possible to find 49 protein species; the difference was generated thanks to the use of the three combined CPLLs. The aberrant behaviour of some proteins, upon treatment via the CPLL method, such as proteins that do not interact with the library, is discussed and evaluated. Simplified elution protocols from the CPLL beads are taken into consideration, of which direct elution in a single step via sodium dodecyl sulphate desorption seems to be quite promising. Alternative methods are suggested. The list of egg yolk components here reported is by far the most comprehensive at present and could serve as a starting point for isolation and functional characterization of proteins possibly having novel pharmaceutical and biomedical applications.
使用包含以伯胺结尾、用末端羧基修饰或用末端叔胺修饰的六肽的组合肽配体文库（CPLL），能够发现和鉴定大量以前未报道的蛋黄蛋白。虽然迄今为止最全面的列表[K. 曼恩、M. 曼恩，《蛋白质组学》，8（2008年）178 - 191]列出了卵黄血浆中约115种独特的基因产物，但我们的发现使这一数值增加了一倍多，达到255种独特的蛋白质种类。从最初未处理的蛋黄中能够找到49种蛋白质种类；这一差异是由于使用了三种组合的CPLL而产生的。文中讨论并评估了一些蛋白质在通过CPLL方法处理时的异常行为，例如不与文库相互作用的蛋白质。考虑了从CPLL磁珠上简化洗脱方案，其中通过十二烷基硫酸钠解吸一步直接洗脱似乎很有前景。还提出了替代方法。此处报告的蛋黄成分列表是目前为止最全面的，可作为可能具有新型药物和生物医学应用的蛋白质分离和功能表征的起点。
