title: The kinetics of conformational fluctuations in an unfolded protein measured by fluorescence methods.
abstract: The simplest dynamic model for an unfolded protein is a statistical coil that continually undergoes substantial conformational fluctuations. A growing number of studies indicate that the unfolded protein is not a simple random coil but rather forms transient structures. We have directly measured the rate of conformational fluctuations of unfolded intestinal fatty acid binding protein (131 aa, 15 kDa) by using fluorescence self-quenching in combination with fluorescence correlation spectroscopy. The conformational fluctuations in this state have an apparent relaxation time, tauR, of 1.6 microsec in 3 M guanidine-HCl at pH 7 and 20 degrees C. The value of tauR increases with increasing solution viscosity, suggesting a diffusive process. In the molten globule state at pH 2, tauR is 2.5 microsec, increasing further with the formation of salt-induced secondary structure. These measurements, which should be widely applicable to other systems, can provide important information about the still incompletely understood conformational properties of unfolded proteins and the mechanism of protein folding.
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, MO 63110, USA.
authors.forename: Krishnananda
authors.initials: K
authors.lastname: Chattopadhyay
authors.forename: Elliot L
authors.initials: EL
authors.lastname: Elson
authors.forename: Carl
authors.initials: C
authors.lastname: Frieden
mesh_terms: D055592:Biophysical Phenomena; D001703:Biophysics; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics; D002942:Circular Dichroism; D050556:Fatty Acid-Binding Proteins; D066298:In Vitro Techniques; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods*; D014783:Viscosity
chemical_list: D002352:Carrier Proteins; D050556:Fatty Acid-Binding Proteins; D011994:Recombinant Proteins