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. To better understand the step-wise changes of the different Trp fluorophores, the spectrum recorded at each temperature was fitted using the discrete states model of Trp residues in proteins (30,32,34)
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. The experimental results indicate that at physiological pH, the dimeric EI^sup SC^ had a well-folded structure; however, at low pH, EI^sup SC^ showed a partially unfolded state with the features of a molten globule, as suggested by fluorescence, far-ultraviolet circular dichroism, FTIR, and 8-anilino-1-naphthalene-sulfonic acid binding
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. The aromatic chromophore 1-anilino-8-naphthalene sulfonate (ANS) is feebly fluorescent in water, but its spectrum is blue shifted and its intensity is dramatically increased in nonpolar solvents or when it binds to nonpolar sites of proteins (4,5; for a general review, see Slavik (6))
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., before the onset of the second calorimetric transition, is fully reversible and leads to no detectable changes in the subsequent DSC scans (data not shown)
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. As an illustration, conformational entropy buildup curves as a function of block size with error bars derived from the block averaging procedure are presented for gp120 atom sets in Fig
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