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. Details about site-directed mutagenesis, protein expression, isolation, and purification of mutants I7S, F110S, and C3A/ C26A have been described elsewhere (22,24)
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. For example, the calculated ratio of k^sub cat^ (DmAChE/HuBChE) for hydrolysis of various substrates at 25°C is 1
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. (20), R^sub 2^ is measured for two settings of δ (we used 2δ^sub 1^ = 450 µs and 2δ^sub 2^ = 3
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. In comparison, the misfolded aggregated states from A-state TTR show a much stronger quantum yield for Bis-ANS than for ANS (Fig
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. The same is true for DCVJ (Fig
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. Glass transition temperature (T^sub g^) and melting temperature (T^sub m^) were determined by DSC (Seiko SII DSC 200C, Perkin Elmer Pyris I DSC)
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. Extensive studies relating to the mechanism of glycosylation and its role in protein folding, stability, and function have been carried out by researchers in the field (7-9)
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. Implications for denatured-state ensembles in strongly denaturing environments Our results have direct bearing on the development of accurate reference-state descriptions for highly denatured proteins
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. When ANS binds to solvent-exposed hydrophobia patches, its quantum yield is enhanced and the maximum of emission is shifted from 520 nm to 480 nm (39,40)
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. The secondary structure of EI^sup sc^ upon binding was not largely affected (Table 4), but there were subtle changes in the stability of the protein (Table 3)
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. In the last decade, strong binding of ANS to molten globule states of proteins has been linked to the loss of tertiary structure (7), and since then the method has been applied to characterize transient states in protein denaturation (8-11)
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. Cytoplasmic concentrations relevant to macromolecular crowding are typically up to 40% v/v (72)
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. Using polyethylene glycol solutions as a model of a crowded cell environment, we studied DNA compaction by alkali metal salts LiCl, NaCl, KCl, RbCl, and CsCl, and found that all of these monocations promote DNA compaction
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.2 M) into a globule (Fig
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. semifasciata, and the vertical bars appeared earlier in the development (seven vertical bars were present in ca
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. We also thank Atila Gosztonyi, Raul Gonzalez, and two anonymous reviewers for providing useful comments on the manuscript
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.7 kJ K^sup -1^ mol^sup -1^) than those in the free form (solid lines until 8 ns)
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. Pyruvate (1 mM), dexamethasone (10^sup -7^ M) and 10 ng/ml TGF-[beta]1 (recombinant human, R&D Systems, Abingdon, UK) were also added
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