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. Structural rearrangements then take place at the dimer interface (and presumably also in other domains of each subunit), leading to a final, compact dimer structure
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. Based on a two-component mixture theory, we obtained the following hydration numbers per protein molecule: 590 +/- 65 for N, 630 +/- 73 for A, and 1110 +/- 67 for U^sub A^
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. This value (311 ± 5 K, at 4 µM protein concentration, 315 ± 9 K, at 6 µM protein concentration) was similar, within the experimental uncertainty, to that obtained in the intact EI^sup ec^ (4), where two tryptophan residues are located at the C-terminal region of the protein
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. These azurins maintain an intact native fold (x-ray structure) but are ~6 kcal/mole less stable relative to the wild-type (WT) (18-22)
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. Upon freezing the sample, the ANS intensity first decreases in the low concentration range, below 80 µM, but then rises sharply above this threshold
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. Since the diffusion is always maintained in some low-dimensional subspace of the full folding space, τ ~ R^sup 2^/D for each subspace, so that τ^sub fold^ ≈ d^sup 2^R^sup 2^/D rather than the exponentially larger (R^sup 2^/R^sup 2^^sub N^)^sup d^
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. The reference solutions corresponding to N, A, and UA states were prepared with HCl into water as those pH values become equal to 5
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. at both excitation wavelengths, showed a bell-shaped profile, with a maximum at neutral pH (Fig
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. These data suggest that the percentages of helical structure in EI^sup sc^ at the extremes of pH were increased
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. This value (311 ± 5 K, at 4 µM protein concentration, 315 ± 9 K, at 6 µM protein concentration) was similar, within the experimental uncertainty, to that obtained in the intact EI^sup ec^ (4), where two tryptophan residues are located at the C-terminal region of the protein
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. The scattered signals were corrected for container scattering, absorption, and instrumental resolution (determined by separate runs of the empty container and of a vanadium sample) by a standard FORTRAN 77 program Iq0 based on the correction formula of Paalman-Pings coefficients (42)
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