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. Although phase diagrams have usually been used in the analysis of Trp fluorescence data in previous publications, the theoretical analysis of the phase diagram is also expected to be true for IR data
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. Similar results were obtained for all other two-state protein sequences shown in Table 1
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. The unite probability of linding large cavities within the interior of denatured proteins emphasizes two points: First, the volume occupied by a chain is filled inefficiently when compared to either a folded protein or a freely rotating chain
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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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. This investigation aims to complement that initial study by addressing the following issues: i), establish whether the perturbation of protein structure reported for azurin is general to other proteins; ii), inquire on potential correlations between the susceptibility to the ice-induced perturbation and specific features of the macromolecule such as thermodynamic stability, internal flexibility, internal cavities/hydration, and covalent links between strands; and finally iii), test for a specific vulnerability of multimeric proteins to freeze damage that may be linked to the disruption of quaternary structure
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. The imaging parameters were as follows: field of view = 2
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. MATERIALS AND METHODSEnzyme purification and characterizationHuBChE was purified to homogeneity from outdated human plasma according to an improved protocol based on that of Grunwald et al
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. To isolate the contribution of the helical junctions to tertiary folding, we also studied the corresponding ribozyme constructs without the loop on arm A (SL 2WJ, 3WJ, and 4WJ, Fig
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. A second major type of error was failure to price the drug at the lowest available price
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