蛋白质相互作用研究的新技术与新方法

目前，蛋白质相互作用已成为蛋白质组学研究的热点. 新方法的建立及对已有技术的改进标志着蛋白质相互作用研究的不断发展和完善．在技术改进方面，本文介绍了弥补酵母双杂交的蛋白定位受限等缺陷的细菌双杂交系统；根据目标蛋白特性设计和修饰TAP标签来满足复合体研究要求的串联亲和纯化技术，以及在双分子荧光互补基础上发展的动态检测多个蛋白质间瞬时、弱相互作用的多分子荧光互补技术．还综述了近两年建立的新方法：与免疫共沉淀相比，寡沉淀技术直接研究具有活性的蛋白质复合体；减量式定量免疫沉淀方法排除了蛋白质复合体中非特异性相互作用的干扰；原位操作的多表位－配基绘图法避免了样品间差异的影响，以及利用多点吸附和交联加固研究弱蛋白质相互作用的固相蛋白质组学方法.

Novel Techniques and Approaches in Protein Protein Interaction Studies

With the fulfillment of human genomic project, proteomics is getting hot. Protein protein interaction studies have become the focus of proteomics. Novel approaches and improved common methods showed the development and complement of proteome research. The article introduced three technique improvements. The bacterial two--hybrid system compensates for limitation of yeast two-hybrid, such as the location of target protein. Novel tags of tandem affinity purification are designed and modified to investigate especial protein complexes, and multicolor fluorescence complementation which is based on bimolecular fluorescence complementation is used to dynamically detect transient and weak interactions among several proteins. New approaches are also summarized. Compared with immunoprecipitation, oligoprecipitation only captures the active form of protein complexes. Quantitative immunoprecipitation combined with knockdown (QUICK) discriminates between specific and nonspecific interactions with an isolated protein complex. Multiepitope-ligand cartography is capable of mapping the location of tens of proteins within the same cell or tissue sample in situ and eliminating errors of different samples. Weak protein-protein interactions are obtained by multiple-site adsorption and cross linked reinforcement in solid phase proteomics strategy.