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. The concentration of ANS, as determined by absorbance at 370 nm, using ε^sub 370^ = 6800 M^sup -1^Cm^sup -1^ (in methanol), ranged between 10 and 450 µM although in most experiments the upper limit was 320 µM
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. The fluorescence emission, collected at 90° from the excitation, is dispersed by a 0
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. Similar plots with relatively lower resolution could be obtained when using me original spectra for the 2D correlation analysis
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. Nevertheless, the 2D IR analysis also suggested that the pretransitional conformational changes involved the unfolding of the solvent-exposed β-sheet structures and aggregates appearing at a temperature at which the protein had not fully unfolded
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. ACYI contains eight Trp residues in each of its two subunits (47,48), and the eight Trp residues are distributed in both the peptidase and dimerization domain (13,17)
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