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Cooperativity induced by a single mutation at the subunit interface of a dimeric enzyme: glutathione reductase
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Details with regard to the intrinsic fluorescence experiments were the same as those described before (30)
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It was suggested, with the help of some computational studies, that this enhanced stability of the tetramer is mainly due to glycosylation and some stabilizing ionic interactions at the oligomeric interface (18)
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The intensities of the bands at different times were measured by densitometry
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The aromatic chromophore 1-anilino-8-naphthalene sulfonate (ANS) is feebly fluorescent in water, but its spectrum is blue shifted and its intensity is dramatically increased in nonpolar solvents or when it binds to nonpolar sites of proteins (4,5; for a general review, see Slavik (6))
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2 and show that the limiting anisotropy level is almost reached after 2 h, which is before the appearance of very large aggregates such as protofibrils and spheres, but is at the peak of the 300-500 kD oligomers
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It is clearer in experiment than the subsequent dimensional reductions because it is the only one that makes significant changes to the radius of gyration of the protein
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We calculated cooling rates as the slope of the average temperature between the top and bottom of a solution versus time at the cloud point of the 10 wt% solution (Figure 2)
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