Department of Physics, North Carolina State University, Raleigh, North Carolina, USA.
Nat Methods. 2010 Mar;7(3):203-5. doi: 10.1038/nmeth.1421. Epub 2010 Jan 31.
We combined single-molecule fluorescence resonance energy transfer (smFRET) with single-particle tracking in live cells to detect the in vivo conformation of individual proteins. We site-specifically labeled recombinant SNARE proteins with a FRET donor and acceptor before microinjecting them into cultured cells. Individual proteins rapidly incorporated into folded complexes at the cell membrane, demonstrating the potential of this method to reveal dynamic interactions within cells.
我们将单分子荧光共振能量转移（smFRET）与活细胞中的单个粒子追踪相结合，以检测单个蛋白质的体内构象。在将重组 SNARE 蛋白微注射到培养的细胞中之前，我们通过在其特定位点标记供体和受体来进行标记。单个蛋白质迅速在细胞膜上组装成折叠复合物，这表明该方法有可能揭示细胞内的动态相互作用。
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