Westbrook Jules A, Wheeler Jun X, Wait Robin, Welson Sandy Y, Dunn Michael J
National Heart & Lung Institute, Imperial College School of Medicine, Heart Science Centre, Harefield Hospital, Harefield, Middlesex, UK.
Electrophoresis. 2006 Apr;27(8):1547-55. doi: 10.1002/elps.200500777.
The analysis of complex proteomes is undertaken using a variety of techniques and technologies such as 2-DE, surface-enhanced laser desorption ionisation, and various types of MS. In order to overcome the complexities of protein expression in discrete proteomes, sample fractionation has become an important aspect of proteomic experiments. The use of narrow-range IPGs (nrIPGs) is of special importance using the 2-DE proteomics workflow, since an enhanced visualisation of a given proteome is achieved through an improved physical separation and resolution of proteins. The work described in this paper presents a series of protein maps of the human heart left ventricle proteome that have been generated using nrIPGs for the first, IEF, dimension of 2-DE. A total of 374 gel spots were excised from seven different pH gradients, covering the range pH 3-10, giving rise to a total of 388 identifications from 110 unique proteins. Using Gene Ontologies (GOs), the identified proteins were found to be associated with 97 types of GO Process, 144 types of GO Function, and 54 types of GO Component. It is hoped that the maps presented in this paper will be of use to other researchers for reference purposes.
复杂蛋白质组的分析采用了多种技术，如二维电泳（2-DE）、表面增强激光解吸电离以及各种类型的质谱（MS）。为了克服离散蛋白质组中蛋白质表达的复杂性，样品分级分离已成为蛋白质组学实验的一个重要方面。在二维电泳蛋白质组学工作流程中，使用窄范围固定化pH梯度（nrIPGs）尤为重要，因为通过改善蛋白质的物理分离和分辨率，可以增强对给定蛋白质组的可视化。本文所述工作展示了一系列人类心脏左心室蛋白质组的蛋白质图谱，这些图谱是首次使用nrIPGs进行二维电泳的第一向等电聚焦（IEF）产生的。从七个不同的pH梯度（覆盖pH 3 - 10范围）中总共切下了374个凝胶点，共鉴定出110种独特蛋白质中的388个。利用基因本体（GO），发现所鉴定的蛋白质与97种GO过程、144种GO功能和54种GO组件相关。希望本文展示的图谱能供其他研究人员参考使用。
