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Detecting the conformation of individual proteins in live cells.

ABSTRACT: 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.

SUBMITTER: Sakon JJ

PROVIDER: S-EPMC2844853 | biostudies-literature | 2010 Mar

REPOSITORIES: biostudies-literature

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Detecting the conformation of individual proteins in live cells.

Sakon John J JJ Weninger Keith R KR

Nature methods 20100131 3

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. ...[more]

PMID: 20118931

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