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. The modeled structure of the EIC^sup sc^ monomer differed from that of EIC^sup tt^ (11) only in a deletion in the loop between α-helix 4 and β-strand 5, which is part of neither the active center nor the dimerization surface
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. 1 shows the change in ANS fluorescence induced by the four azurin species upon freezing of the sample at -13°C (200 µM ANS, 2
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. Between 2 and 3 M GdnHCl, two partially folded intermediate states possessing flexible tertiary structures (revealed by fluorescence data) but compact secondary structures (as indicated by circular dichroism measurements) were identified
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. since at this pH the fluorescence intensity was maximal (Fig
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. In contrast, the slowest heating rate of 0
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. Part of this uncertainty arises as a result of the polyampholyte characteristics of the protein
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. In addition to the divergence of viscosity (Fig
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. In this method, the fluorescence of donor and acceptor are simultaneously recorded using avalanche-photodiodes as individual molecules traverse the excitation volume in a confocal microscope, and the FRET efficiencies from many such single-molecule observations are plotted in the form of a histogram
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