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. 1 B), whereas iodide quenching and CD measurements revealed significant preservation of GR structure at 3 M GdnHCl
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. Detection and characterization of the intermediate on the folding pathway of human alpha-lactalbumin
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. Although gSBA was seen to demonstrate very high conformational stability, rSBA exhibits comparatively much lower firmness to resist the structural perturbation induced by the denaturant guanidinium hydrochloride
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. The helix-loop-helix motif formed by the α-helix 9-α-helix 10 shows intermonomer salt bridges: Asp^sup 540^ in the motif with Arg^sup 422^' in α-helix 5 from the other monomer (the prime is used to designate residues in the other monomer)
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. Rather solvent accessible tryptophan residues in the folded state (Fig
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. In an earlier NMR study we could conclude that the CaM:SEF2-1mp complex has a 2:2 stoichiometry, where two interacting CaM molecules bind one homodimeric SEF2-1mp (11)
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. Still the formed aggregates were too large to enable direct determination of their rotational correlation time due to the modest lifetime of ANS although being 16 ns
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. The cooling rate average between the top and bottom of the solution near the cloud point (approximately 100°C, determined by visual observation) was approximately 11°C/min
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. We examined a series of six different ribozymes consisting of two-, three- and four-way junction variants, as well as corresponding constructs with one of the two loops removed
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. The quasi-docked conformation interconverts rapidly with an extended structure, giving rise to continuous peak shifts with added Mg^sup 2+^, as well as pronounced asymmetry and broadening in the FRET efficiency histogram at high Mg^sup 2+^ concentrations
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. Thus, to record adequate emission spectra for GR, we routinely started acquisition of the spectra at 295 nm (as shown in Fig
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. coli GR: comparison of human and yeast GR yields ~68% of similarity and ~52% of sequence identity between the two proteins
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. The role of glycosylation in relation to protein structure, form, and stability has been a topic of much interest for a long time
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. To derive thermodynamic parameter from spectroscopic data it is crucial whether the signal changes during the unfolding process are proportional to the molar fractions of the folded and unfolded protein
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.Mikael Lindgren,* Karin Sörgjerd,[dagger] and Per Hammarström[dagger]* Department of Physics, Norwegian University of Science and Technology, 7491 Trondheim, Norway; and [dagger] IFM-Department of Chemistry, Linköping University, SE-581 83 Linköping, SwedenSubmitted July 16, 2004, and accepted for publication March 9, 2005
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. This apparent K^sub d^, which reflects protein-protein associations on a confined membrane surface, is predicted to be ~2 orders of magnitude weaker when the proteins are free to associate in solution (see Runnels and Scarlata, 1999)
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.We repeated this study in the absence of membranes using freshly dialyzed CPM-PLCβ-Gβγ and found that in solution the volume change increases to 78 ± 3 ml/mol
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. Disk formation was confirmed by negative staining electron microscopy (EM) and non-denaturing gel electrophoresis which showed no free protein
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