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The precipitate was resuspended m 6 M guanidinium hydrochloride (in 10 mM Tris buffer (pH 7
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. The signal was acquired every second, and the increment of wavelength was set to 1 nm
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. For example, more compact structures of the thermal unfolded states as compared to chemical unfolded states would also explain more buried tryptophan residues which exhibit smaller red shifts
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. The newly frozen samples were then allowed to anneal for 12 h at -5°C, followed by a final equilibration of 2 h at the temperature selected for fluorescence measurements (-5 and -13°C)
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- Phenylalanine side chain behavior of the intestinal fatty acid-binding protein: the effect of urea on backbone and side chain stability
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. Taken together, these data suggest that there were structural changes, which partially solvent-exposed the indole moiety and/or tyrosine residues, and modified the ellipticity at 222 nm (41,42)
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. Cell Death Differ 2004; 11:1258-1264(41) Morimoto K, Amano H, Sonoda F, et al
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. This shoulder seems to represent a subpopulation of thermal unfolded states which exhibit less buried tryptophan residues, possibly related to unfolded proteins which are not aggregated
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