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. ANS binding experiments, extended to a set of proteins including serum albumin, α-amylase, β-galactosidase, alcohol dehydrogenase from horse liver, alcohol dehydrogenase from yeast, lactic dehydrogenase, and aldolase, confirmed that a stressed condition of the native fold in the frozen state appears to be general to most proteins and pointed out that oligomers tend to be more labile than monomers presumably because the globular fold can be further destabilized by subunit dissociation
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. In agreement with the previous report (12) for WT azurin, the intensity is enhanced ~7-fold and the spectrum is blue shifted by 13 nm relative to the ANS control
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. Mass normalized fluorescence profiles are shown in Fig
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. The column was then washed extensively with PBS till the washings had A^sub 280^Isothermal guanidinium-hydrochloride-induced denaturation Equilibrium unfolding as a function of guanidinium hydrochloride (GdnCl) concentration was monitored by fluorescence spectroscopy
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. In conclusion, our approach suggests that thermodynamical analysis might reveal subtle contormational changes in otherwise similar folds
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.33 K/min is observed above 80°C, ~10°C higher than the onset of the corresponding CD transition
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