Institut de Biotecnologia i de Biomedicina, Departament de Bioquímica i Biologia Molecular, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.
Curr Mol Med. 2011 Oct;11(7):582-98. doi: 10.2174/156652411800615117.
Many cellular processes depend on the establishment of selective stable or transient interactions between proteins. Therefore, the ability to identify and characterize these contacts in physiologically relevant environments is crucial to understanding the networks of contacts that allow the transmission and integration of biological information in living cells. Protein-fragment complementation assays (PCA) have emerged as approaches that report on the proximity of two given proteins in the cell at a given location and time. In particular, bimolecular fluorescence complementation (BIFC) allows noninvasive imaging of protein binding in living cells at high spatial resolution and without the requirement for exogenous substrates. In the present review, we discuss PCA and BIFC fundamentals, the implementation of BIFC assays and selected applications of BIFC in drug discovery, developmental studies or neurological disorders.
许多细胞过程依赖于蛋白质之间选择性稳定或瞬时相互作用的建立。因此，能够在生理相关环境中识别和描述这些接触点对于理解允许生物信息在活细胞中传递和整合的接触网络至关重要。蛋白片段互补测定（PCA）已经成为一种方法，可以报告在特定位置和时间下两个给定蛋白在细胞中的接近程度。特别是，双分子荧光互补（BIFC）允许在高空间分辨率下非侵入性地对活细胞中的蛋白质结合进行成像，而无需外源底物。在本综述中，我们讨论了 PCA 和 BIFC 的基本原理、BIFC 测定的实施以及 BIFC 在药物发现、发育研究或神经紊乱中的应用。
