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To assay conformational preferences of highly denatured proteins, we quantify a variety of properties for EV-limit ensembles of 23 two-state proteins
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, (48)), thus compensating for the reduced flexibility adjacent to the binding site
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5 ns with squared order parameter (S^sup 2^) of ~0
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Literature CitedAigner, P
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American Midland Naturalist 136:149-156
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Fong R, Kissmeyer-Nielsen F
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. Address reprint requests to Makoto Suzuki, Department of Metallurgy, Graduate School of Engineering, Tohoku University, Aoba-yama 02, Sendai, 980-8579, Japan
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. Interestingly, the results from fluorescence and infrared (IR) spectroscopy clearly indicated that a pretransitional stage existed at temperatures from 50°C to 66°C
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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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. Inasmuch as these theories are applicable to denatured proteins, the current work shows that many of the seemingly paradoxical observations regarding denatured proteins are readily resolved
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. Protein was extensively dialyzed against 2 L of the working buffer (5 mM HEPES buffer containing 10 mM NaCl and 2 mM β-mercaptoethanol, pH 7) at 278 K
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. Lyophilized BSA, β-gal from Aspergillus oryzae and yeast alcohol dehydrogenase (YADH), and crystalline suspensions of α-amylase from porcine pancreas, ALD from rabbit muscle and LDH from rabbit muscle (LDH) were purchased from Sigma-Aldrich Co
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. The newly frozen samples were then allowed to anneal for 12 h at -5°C, followed by a final equilibration of 2 h at the temperature selected for fluorescence measurements (-5 and -13°C)
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. Both lyophilized powders were placed in aluminum sample holders (30 × 40 × 0
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. Early measurements (42) showed BLG has a dipole moment of a sufficient magnitude to generate significant attractive dipole-dipole interactions which are predicted to be screened by increasing ionic strength at the concentrations relevant to this study (40,43)
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. The effect of temperature and ionic strength on the dimerisation of beta-lactoglobulin
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