Ciruela Francisco
Unitat de Farmacologia, Departament de Patologia i Terapèutica Experimental, Facultat de Medicina (Campus de Bellvitge), IDIBELL-Universitat de Barcelona, 08907 L'Hospitalet del Llobregat, Barcelona, Spain.
Curr Opin Biotechnol. 2008 Aug;19(4):338-43. doi: 10.1016/j.copbio.2008.06.003. Epub 2008 Jul 16.
Multiprotein complexes partake in nearly all cell functions, thus the characterization and visualization of protein-protein interactions in living cells constitute an important step in the study of a large array of cellular mechanisms. Recently, noninvasive fluorescence-based methods using resonance energy transfer (RET), namely bioluminescence-RET (BRET) and fluorescence-RET (FRET), and those centered on protein fragment complementation, such as bimolecular fluorescence complementation (BiFC), have been successfully used in the study of protein interactions. These new technologies are nowadays the most powerful approaches for visualizing the interactions occurring within protein complexes in living cells, thus enabling the investigation of protein behavior in their normal milieu. Here we address the individual strengths and weaknesses of these methods when applied to the study of protein-protein interactions.
多蛋白复合物几乎参与所有细胞功能，因此活细胞中蛋白质 - 蛋白质相互作用的表征和可视化是研究大量细胞机制的重要一步。最近，利用共振能量转移（RET）的非侵入性荧光方法，即生物发光共振能量转移（BRET）和荧光共振能量转移（FRET），以及以蛋白质片段互补为中心的方法，如双分子荧光互补（BiFC），已成功用于蛋白质相互作用的研究。如今，这些新技术是可视化活细胞中蛋白质复合物内发生的相互作用的最强大方法，从而能够在其正常环境中研究蛋白质行为。在此，我们阐述这些方法在应用于蛋白质 - 蛋白质相互作用研究时各自的优缺点。
