Yano Yoshiaki, Matsuzaki Katsumi
Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, Japan.
Biochim Biophys Acta. 2009 Oct;1788(10):2124-31. doi: 10.1016/j.bbamem.2009.07.017. Epub 2009 Jul 30.
Instead of using reconstituted proteoliposomes, in situ investigations of membrane proteins in living cell membranes are important because the heterogeneous and dynamic nature of biomembranes significantly affects their behavior. Protein-specific labeling is a key technique for the detection of a target protein by fluorescence measurements, particularly fluorescence microscopy. However, conventional genetic fusion with fluorescent proteins has several shortcomings. Post-translational labeling methods using a genetically encodable tag and synthetic probes targeting to the tag can overcome these limitations. This review summarizes emerging tag-probe techniques for labeling specific membrane proteins and their applications, including endocytotic internalization, partitioning to specific membrane domains, interprotein interactions, and conformational changes.
相比于使用重组蛋白脂质体，对活细胞膜中的膜蛋白进行原位研究很重要，因为生物膜的异质性和动态性质会显著影响其行为。蛋白质特异性标记是通过荧光测量，特别是荧光显微镜检测目标蛋白的关键技术。然而，与荧光蛋白的传统基因融合存在几个缺点。使用可遗传编码标签和靶向该标签的合成探针的翻译后标记方法可以克服这些限制。本文综述了用于标记特定膜蛋白的新兴标签-探针技术及其应用，包括内吞内化、分配到特定膜结构域、蛋白间相互作用和构象变化。
