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Single Molecule FRET of Protein Folding Dynamics
Author(s) -
Schuler Ben
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.327.1
Subject(s) - förster resonance energy transfer , chemistry , chemical physics , protein folding , folding (dsp implementation) , single molecule experiment , intramolecular force , molecule , crystallography , biophysics , context (archaeology) , downhill folding , energy landscape , native state , unfolded protein response , single molecule fret , phi value analysis , molecular dynamics , fluorescence , computational chemistry , physics , stereochemistry , biochemistry , quantum mechanics , electrical engineering , biology , engineering , paleontology , organic chemistry , endoplasmic reticulum
We have been using single molecule Förster resonance energy transfer (FRET) for the investigation of protein folding, with a focus on non‐native states of proteins; these are conformationally more heterogeneous than native structures and have thus been more difficult to access experimentally. Single molecule FRET has allowed us to map long‐range distance distributions in the unfolded state of the small cold shock protein Csp Tm 1 , even at low concentrations of denaturant, where the unfolded molecules are collapsed, and where the majority of protein molecules are already folded. Using advanced correlation methods, we determined the reconfiguration time of unfolded Csp Tm 2 and found a decrease in its intramolecular diffusion coefficient upon collapse. This increase in the roughness of the free energy surface is accompanied by the formation of some secondary structure, as determined with kinetic synchrotron circular dichroism experiments 1 . Complete fluorescence intensity correlation functions from picoseconds to seconds are an increasingly important tool to investigate protein dynamics 3 and quantify previously inaccessible characteristics of the free energy surfaces for folding. In a next step, the same methods now allow us to investigate the folding and dynamics of proteins in the context of cellular factors, such as molecular chaperones. 4