TY  - JOUR
ID  - open770
UR  - http://www.sciencedirect.com/science/article/pii/S0301462296022442
IS  - 2-3
A1  - Guptasarma, Purnananda
N2  - Unlike steady-state spectrofluorimetry, time-resolved emission spectroscopy (TRES) can resolve emissions from fluorophores with similar quantum yields and overlapping steady-state emission spectra. Time-resolved emission studies of the protein-intrinsic fluorophore, tryptophan (Trp), can thus potentially be used to examine protein conformational heterogeneity in solution, as well as to investigate the existence of populated intermediate structural states in equilibrium unfolding reactions of single-tryptophan proteins. Here, the single-Trp copper protein, azurin, is examined in various concentrations of guanidine hydrochloride (GdnCl) with its disulphide bond in an intact state. Interestingly, multiple envelopes of Trp emission are observed in all TRES spectra acquired, instead of just two emission envelopes (corresponding to the native and unfolded states) expected from two-state unfolding. These envelopes appear to be centred around the same set of emission wavelengths in different TRES spectra, and only intensities and decay rates vary with the concentration of denaturant used. This suggests that structural states representing different levels of exposure of Trp to the aqueous solvent might, in fact, be populated at equilibrium during the unfolding of azurin by GdnCl.
VL  - 65
TI  - Resolving multiple protein conformers in equilibrium unfolding reactions: a time-resolved emission spectroscopic (TRES) study of Azurin.
AV  - restricted
EP  - 8
N1  - Copyright of this article belongs to Elsevier Science.
Y1  - 1997/04/22/
PB  - Elsevier Science
JF  - Biophysical chemistry
KW  - Fluorescence decay time-resolved emission; Protein conformation; Protein folding; Structural intermediates
SN  - 0301-4622
SP  - 221
ER  -