Schmidt Antje, Wiesner Burkhard, Schülein Ralf, Teichmann Anke
Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Straße 10, 13125, Berlin, Germany.
Methods Mol Biol. 2014;1174:139-56. doi: 10.1007/978-1-4939-0944-5_9.
The fusion of fluorescent proteins to G protein-coupled receptors (GPCRs) is an important tool to study, e.g., trafficking and protein interactions of these important drug targets. In the past, the green fluorescent protein and its derivatives have been widely used as fluorescent tags. More recently, it was reported that photoconvertible fluorescent proteins (PCFPs) such as Kaede or Kikume green-red protein could also be used as fluorescent tags for GPCRs. These proteins have the obvious advantage that their fluorescence can be switched once the GPCR of interest has reached a specific subcellular compartment. Here, we summarize the recent progress for live cell imaging of GPCRs using these PCFPs for trafficking, biosynthesis, and protein/protein interaction studies.
将荧光蛋白与G蛋白偶联受体（GPCRs）融合是研究这些重要药物靶点的运输和蛋白质相互作用等的重要工具。过去，绿色荧光蛋白及其衍生物被广泛用作荧光标签。最近，有报道称可光转换荧光蛋白（PCFPs），如Kaede或Kikume绿-红蛋白，也可作为GPCRs的荧光标签。这些蛋白具有明显的优势，即一旦感兴趣的GPCR到达特定的亚细胞区室，其荧光就可以被切换。在此，我们总结了使用这些PCFPs进行GPCRs活细胞成像以用于运输、生物合成和蛋白质/蛋白质相互作用研究的最新进展。
