膜转运蛋白的功能性超表达和纯化

膜转运蛋白结构和功能的研究是功能膜蛋白质组研究中的一个重要内容，而大量蛋白质的分离纯化是进行蛋白质的结构和功能研究的基础.目前,结构和功能膜蛋白质组学相关研究的瓶颈,在于不能有效地超量表达和纯化具有生物活性的膜转运蛋白.影响膜转运蛋白超量表达和纯化的关键因素,包括目标蛋白的拓扑学结构分析和去垢剂的选择.进行膜转运蛋白拓扑学结构的分析,对于构建用于活体表达的重组膜转运蛋白具有指导意义.去垢剂能够稳定去膜状态的膜蛋白，在膜转运蛋白的离体表达和亲和纯化以及包涵体的处理过程中具有重要的作用.本文就目前功能膜蛋白质组学研究中所涉及的有关膜转运蛋白功能性超表达和分离纯化策略及关键技术作一简述.

Overexpression and Purification of Membrane Transport Protein with Native Functions

Structural and functional characterization of membrane transport protein is very important in functional membrane proteomics, which relies on the isolation and purification of large amount of functional proteins. However, effective overexpression and purification of biologically active membrane transport proteins are the primary bottleneck in contemporary functional and structural proteomics studies. Crucial factors influencing membrane protein overproductions include topological analysis of target proteins and selections of detergents. Topological analysis provides valuable information in constructing recombinant membrane transport proteins for in vivo expression. Detergents, which used to maintain the native structure and function of lipid-free membrane proteins, play important roles in cell-free expression, affinity purification of membrane transport proteins, and in vitro their refolding from inclusion bodies. In this article, we reviewed the key strategies and crucial techniques for the overexpression and purification of membrane transport proteins in functional proteomics.