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The dialyzed solution was centrifuged at 8000 rpm for 15 min and the clear supernatant was loaded on a lactosylamine Biogel P-150 column preequilibrated wilh buffer (21)
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We thank Maria Francisca Lozano for her involvement in the early stages of this work and Francisco N
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We note that the term "compact" is used in this article to denote the proximity of arms A and B, as described by the FRET efficiency for the current labeling scheme
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- We could then discuss the relation between structure change and hydration of protein, together with other thermodynamic parameters
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. In experiments aimed at detecting the burial of the aromatic residues upon binding to PEP and/or Mg^sup 2+^, the concentration of EI^sup sc^ was 4 µM at pH 7
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. Thermal denaturation experiments were also carried out by following the emission fluorescence of ANS either at 480 or 520 nm, after excitation at 360, 380, or 390 nm, at three different pH, 2
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. In conclusion, residues with fast exchange can be identified from the combination of R^sub ex^ and ΔR^sub ex^ using either the fast exchange equations or the general fast/slow exchange equations derived by Ishima and Torchia (34)
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. It should be noted that DCVJ reaches the plateau level faster than ThT and ANS, indicating a strong preference for oligomeric aggregates
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.03 kJ K^sup -1^ mol^sup -1^), indicating that the complex form samples slightly different parts of conformational space (Fig
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. The quasi-docked conformation interconverts rapidly with an extended structure, giving rise to continuous peak shifts with added Mg^sup 2+^, as well as pronounced asymmetry and broadening in the FRET efficiency histogram at high Mg^sup 2+^ concentrations
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