Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, UK.
Biochem Soc Trans. 2010 Aug;38(4):875-8. doi: 10.1042/BST0380875.
In the present article, we describe the two standard high-throughput methods for identification of protein complexes: two-hybrid screens and TAP (tandem affinity purification) tagging. These methods have been used to characterize the interactome of Saccharomyces cerevisiae, showing that the majority of proteins are part of complexes, and that complexes typically consist of a core to which are bound 'party' and 'dater' proteins. Complexes typically are merely the sum of their parts. A particularly interesting type of complex is the metabolon, containing enzymes within the same metabolic pathway. There is reasonably good evidence that metabolons exist, but they have not been detected using high-thoughput assays, possibly because of their fragility.
在本文中，我们描述了两种鉴定蛋白质复合物的标准高通量方法：双杂交筛选和 TAP（串联亲和纯化）标记。这些方法已被用于表征酿酒酵母的相互作用组，表明大多数蛋白质都是复合物的一部分，并且复合物通常由核心组成，核心结合有“伙伴”和“约会”蛋白。复合物通常仅仅是其组成部分的总和。一种特别有趣的复合物类型是代谢物，其中包含同一代谢途径中的酶。有相当充分的证据表明代谢物的存在，但它们没有通过高通量检测来检测到，可能是因为它们不稳定。
