Ontario Cancer Institute, University Health Network, Canada.
Expert Rev Proteomics. 2010 Feb;7(1):141-54. doi: 10.1586/epr.09.97.
Defining the cell surface proteome has profound importance for understanding cell differentiation and cell-cell interactions, as well as numerous pathogenic abnormalities. Owing to their hydrophobic nature, plasma membrane proteins that reside on the cell surface pose analytical challenges and, despite efforts to overcome difficulties, remain under-represented in proteomic studies. Limitations in the classically employed ultracentrifugation-based approaches have led to the invention of more elaborate techniques for the purification of cell surface proteins. Three of these methods--cell surface coating with cationic colloidal silica beads, biotinylation and chemical capture of surface glycoproteins--allow for marked enrichment of this subcellular proteome, with each approach offering unique advantages and characteristics for different experiments. In this article, we introduce the principles of each purification method and discuss applications from the recent literature.
定义细胞表面蛋白质组对于理解细胞分化和细胞-细胞相互作用以及许多致病异常具有重要意义。由于其疏水性，位于细胞表面的质膜蛋白存在分析挑战，尽管已经努力克服困难，但在蛋白质组学研究中仍未得到充分体现。经典的基于超速离心的方法的局限性导致了更复杂的细胞表面蛋白纯化技术的发明。其中三种方法——阳离子胶体硅珠对细胞表面的涂层、生物素化和表面糖蛋白的化学捕获——可以显著富集这个亚细胞蛋白质组，每种方法在不同的实验中都有独特的优势和特点。在本文中，我们介绍了每种纯化方法的原理，并讨论了来自最近文献的应用。
