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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 cu...

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Detalles Bibliográficos
Autores principales: Sakon, John J., Weninger, Keith R.
Formato: Texto
Lenguaje:English
Publicado: 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2844853/
https://www.ncbi.nlm.nih.gov/pubmed/20118931
http://dx.doi.org/10.1038/nmeth.1421
Descripción
Sumario: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. We observed that individual proteins rapidly incorporated into folded complexes at the cell membrane, demonstrating the potential of this method to reveal dynamic interactions within cells.