生物膜是一类重要的亚细胞结构,含有大量的跨膜蛋白和非跨膜蛋白,负责细胞与外界的物质和信息交换以及行使细胞内的多种功能。鉴定并分析膜蛋白在生物膜上的表达是研究其功能必不可少的步骤。蛋白质组学技术可以在单次实验中一次性从全细胞裂解物中鉴定多达上万个蛋白质,从而为分析这些蛋白的表达提供了直接的证据。但由于膜蛋白具有较强的疏水性,从而导致对膜蛋白的蛋白质组学鉴定相对困难,这也使得对膜蛋白的结构与功能的研究进展相对比较缓慢。随着鸟枪法蛋白质组学(shot-gun proteomics)及滤膜辅助的样品制备(filter aided sample preparation)等为代表的现代蛋白质组学技术的快速发展,膜蛋白的鉴定水平及覆盖率也随之大幅提高。本文主要综述了过去20余年在膜蛋白质组学技术上的重要进展,并以光合作用模式蓝藻集胞藻(Synechocystis sp. PCC6803)的膜蛋白质组学研究为例,阐述了技术的进步如何提高膜蛋白鉴定的覆盖度,以期为其他物种膜蛋白质组全覆盖度的鉴定提供相应理论借鉴。

Membrane proteins play important functions not only as receptors and transporters, but also in many other important intracellular functions such as photosynthetic and respiratory electron transport. Identification of membrane proteins is a necessary step to understand their functions. Membrane proteins are generally highly hydrophobic and difficult to be resolved by aqueous solutions, and large-scale proteomic identification of membrane proteins has been a great technical challenge. Significant efforts have been invested in the field to improve the solubility of membrane proteins in aqueous solutions that are compatible for mass spectrometry analysis. This review summarizes the main technological achievements in the field of membrane proteomics particularly for the improvement of membrane protein identification, and uses the photosynthetic model cyanobacterium Synechocystis sp. PCC6803 as an example to illustrate how technology advances push forward the field in terms of the increased coverage of membrane proteome identification.