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The fluorescence emission of GR undergoes a marked red-shift between 1 and 3 M GdnHCl (with a transition midpoint at ~2
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5 C was not reliable due to the lack of some spectra, as mentioned above
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Department of Agriculture, Beltsville Human Nutrition Research Center, Beltsville, Maryland
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2 M^sup -1^ and v = 0
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In either case the conclusion is that in the frozen state the polypeptide structure of the putative binding sites has changed, becoming unable to host ANS or to enhance its fluorescence yield, as when the probe surrounding changes from rigid to mobile (26)
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At each sampling time, the state of the system is queried, and the recorded photon is assigned to either donor or acceptor channel, with a probability based on the FRET efficiency of the detected system state
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3 A for comparison), and these bursts are analyzed to generate a FRET histogram (Fig
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We refer to this species as a quasi-docked state to denote the native-like arrangement of arms A and B, although there are no actual docking interactions between these arms because loop A is absent
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In summary, our comparative study of different ribozyme constructs shows that in contrast with the 2WJ and the 3WJ, the natural 4WJ architecture can efficiently guide conformational search events during ribozyme folding, thereby dramatically influencing subsequent tertiary structure formation and stability
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. By using a combination of fluorescence and circular dichroism spectroscopies, thiol group titration, and size-exclusion chromatography we have detected the existence of partially folded intermediate states in the unfolding of GR
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