Detecting the conformation of individual proteins in live cells.
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.
Sakon
John J
JJ
Department of Physics, North Carolina State University, Raleigh, North Carolina, USA.
Weninger
Keith R
KR
D002478:Cells, Cultured; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D008561:Membrane Fusion; D011487:Protein Conformation; D017510:Protein Folding; D050600:SNARE Proteins / Q000737:chemistry*; D050825:Synaptosomal-Associated Protein 25
D050600:SNARE Proteins; D050825:Synaptosomal-Associated Protein 25