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Three-dimensional molecular modeling with single molecule FRET.


ABSTRACT: Single molecule fluorescence energy transfer experiments enable investigations of macromolecular conformation and folding by the introduction of fluorescent dyes at specific sites in the macromolecule. Multiple such experiments can be performed with different labeling site combinations in order to map complex conformational changes or interactions between multiple molecules. Distances that are derived from such experiments can be used for determination of the fluorophore positions by triangulation. When combined with a known structure of the macromolecule(s) to which the fluorophores are attached, a three-dimensional model of the system can be determined. However, care has to be taken to properly derive distance from fluorescence energy transfer efficiency and to recognize the systematic or random errors for this relationship. Here we review the experimental and computational methods used for three-dimensional modeling based on single molecule fluorescence resonance transfer, and describe recent progress in pushing the limits of this approach to macromolecular complexes.

SUBMITTER: Brunger AT 

PROVIDER: S-EPMC3051805 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature

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Three-dimensional molecular modeling with single molecule FRET.

Brunger Axel T AT   Strop Pavel P   Vrljic Marija M   Chu Steven S   Weninger Keith R KR  

Journal of structural biology 20100917 3


Single molecule fluorescence energy transfer experiments enable investigations of macromolecular conformation and folding by the introduction of fluorescent dyes at specific sites in the macromolecule. Multiple such experiments can be performed with different labeling site combinations in order to map complex conformational changes or interactions between multiple molecules. Distances that are derived from such experiments can be used for determination of the fluorophore positions by triangulati  ...[more]

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