Ivanusic Daniel, Denner Joachim, Bannert Norbert
Robert Koch Institute, HIV and Other Retroviruses, Berlin, Germany.
Curr Protoc Protein Sci. 2016 Aug 1;85:29.17.1-29.17.13. doi: 10.1002/cpps.10.
This unit provides a guide and detailed protocol for studying membrane protein-protein interactions (PPI) using the acceptor-sensitized Förster resonance electron transfer (FRET) method in combination with the proximity ligation assay (PLA). The protocol in this unit is focused on the preparation of FRET-PLA samples and the detection of correlative FRET/PLA signals as well as on the analysis of FRET-PLA data and interpretation of correlative results when using cyan fluorescent protein (CFP) as a FRET donor and yellow fluorescent protein (YFP) as a FRET acceptor. The correlative application of FRET and PLA combines two powerful tools for monitoring PPI, yielding results that are more reliable than with either technique alone. © 2016 by John Wiley & Sons, Inc.
本单元提供了一个指南和详细方案，用于结合邻近连接分析（PLA），使用受体敏化的Förster共振电子转移（FRET）方法研究膜蛋白-蛋白相互作用（PPI）。本单元中的方案重点在于FRET-PLA样品的制备、相关FRET/PLA信号的检测，以及在使用青色荧光蛋白（CFP）作为FRET供体和黄色荧光蛋白（YFP）作为FRET受体时FRET-PLA数据的分析和相关结果的解读。FRET和PLA的相关应用结合了两种强大的监测PPI的工具，所产生的结果比单独使用任何一种技术都更可靠。© 2016约翰威立国际出版公司。
