Protein-Protein Interactions: Co-Immunoprecipitation
Protein-Protein Interactions: Co-Immunoprecipitation
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DOI：
10.1007/978-1-4939-7033-9_17
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发表时间：
2017-01-01
期刊：
影响因子：
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通讯作者：
Lai, Erh-Min
中科院分区：
文献类型：
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作者：
Lin, Jer-Sheng;Lai, Erh-Min
蛋白质通常不是作为单一物质发挥作用，而是作为动态网络中的团队成员。越来越多的证据表明，蛋白质-蛋白质相互作用在活细胞中的许多生物过程中至关重要。遗传学（如酵母双杂交，Y2 H）和生物化学（如免疫共沉淀，co-IP）方法是在研究开始时常用的方法，以确定相互作用的蛋白质。免疫沉淀（IP）是一种使用靶蛋白特异性抗体与蛋白A/G亲和珠结合的方法，是鉴定与特定蛋白相互作用的分子的有力工具。因此，co-IP被认为是鉴定或确认体内蛋白质-蛋白质相互作用事件发生的标准方法之一。Co-IP实验可以通过直接或间接的相互作用或蛋白质复合物来鉴定蛋白质。在这里，我们使用农杆菌VI型分泌系统（T6 SS）鞘组件TssB-TssC（41）的相互作用作为一个例子来描述co-IP的原理，程序和实验问题。
Proteins often do not function as single substances but rather as team players in a dynamic network. Growing evidence shows that protein-protein interactions are crucial in many biological processes in living cells. Genetic (such as yeast two-hybrid, Y2H) and biochemical (such as co-immunoprecipitation, co-IP) methods are the methods commonly used at the beginning of a study to identify the interacting proteins. Immunoprecipitation (IP), a method using a target protein-specific antibody in conjunction with Protein A/G affinity beads, is a powerful tool to identify molecules that interact with specific proteins. Therefore, co-IP is considered to be one of the standard methods of identifying or confirming the occurrence of protein-protein interaction events in vivo. Co-IP experiments can identify proteins via direct or indirect interactions or in a protein complex. Here, we use Agrobacterium type VI secretion system (T6SS) sheath components TssB-TssC(41) interaction as an example to describe the principle, procedure, and experimental problems of co-IP.