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18 is less than 18 full units of charge due to charge regulation-adjustment of protonation states of other ionizable groups on the protein to counteract the increasing electrostatic potential on its surface. See ref 15 for details.
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18
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It is possible that by using fluorescence of DNSA as a marker for the folded state of the protein, we cannot observe small perturbations in the three-dimensional structure, if these perturbations do not affect binding of DNSA. By using DNSA, we also cannot observe intermediates or changes in the secondary structure.
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We observed that DNSA, when present with the protein in the denaturing solution, reduced the rates of denaturation by factors of 2-10, but further studies are necessary to quantify the effect more precisely.
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Manuscript in preparation
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