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10.1021/acs.jpcb.7b03441

http://scihub22266oqcxt.onion/10.1021/acs.jpcb.7b03441
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suck abstract from ncbi


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pmid28709377
      J+Phys+Chem+B 2017 ; 121 (35 ): 8211-8241
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  • Combining Graphical and Analytical Methods with Molecular Simulations To Analyze Time-Resolved FRET Measurements of Labeled Macromolecules Accurately #MMPMID28709377
  • Peulen TO ; Opanasyuk O ; Seidel CAM
  • J Phys Chem B 2017[Sep]; 121 (35 ): 8211-8241 PMID28709377 show ga
  • Förster resonance energy transfer (FRET) measurements from a donor, D, to an acceptor, A, fluorophore are frequently used in vitro and in live cells to reveal information on the structure and dynamics of DA labeled macromolecules. Accurate descriptions of FRET measurements by molecular models are complicated because the fluorophores are usually coupled to the macromolecule via flexible long linkers allowing for diffusional exchange between multiple states with different fluorescence properties caused by distinct environmental quenching, dye mobilities, and variable DA distances. It is often assumed for the analysis of fluorescence intensity decays that DA distances and D quenching are uncorrelated (homogeneous quenching by FRET) and that the exchange between distinct fluorophore states is slow (quasistatic). This allows us to introduce the FRET-induced donor decay, ?(D)(t), a function solely depending on the species fraction distribution of the rate constants of energy transfer by FRET, for a convenient joint analysis of fluorescence decays of FRET and reference samples by integrated graphical and analytical procedures. Additionally, we developed a simulation toolkit to model dye diffusion, fluorescence quenching by the protein surface, and FRET. A benchmark study with simulated fluorescence decays of 500 protein structures demonstrates that the quasistatic homogeneous model works very well and recovers for single conformations the average DA distances with an accuracy of
  • |*Fluorescence Resonance Energy Transfer [MESH]
  • |*Molecular Dynamics Simulation [MESH]
  • |Diffusion [MESH]
  • |Fluorescence [MESH]
  • |Fluorescent Dyes/*chemistry [MESH]
  • |Macromolecular Substances/analysis [MESH]
  • |Proteins/*analysis [MESH]


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