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That all mutant proteins exhibit a comparable modest negative cooperativity is also consistent with a common mode of binding.
That MftR binds urate with modest negative cooperativity is consistent with the existence of two sites; such negative cooperativity of urate binding was also observed for HucR and PecS.
MftR was previously shown to bind two sites in the mftR-mftP intergenic region (mftO; Figure 1) with modest negative cooperativity, and the ligand urate was shown to attenuate DNA binding.
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The resulting signal weighted average sedimentation coefficients are not consistent with models that assume even modest 10-fold positive or negative cooperativity.
Conversely, negative cooperativity makes enzymes insensitive to small changes in [S].
Interestingly, the consistently low Hill slope observed here and previously 13 could indicate negative cooperativity restricting ligand occupancy on all four subunits of the tetramer.
In contrast, only minor changes in equilibrium binding are observed under conditions of strong negative cooperativity (Fig. 1 C).
Consistently, several pairwise interactions within a small subset of proteins, most notably ETS1 and GATA3, showed moderate negative cooperativity in p65 recruitment (Fig. 4a).
In negative cooperativity, the binding of a molecule to the first subunit makes more difficult the binding of substrate to the second.
Biotinylated DNA showed strong negative cooperativity of binding to cis-divalent but not trans-divalent streptavidin.
A concave upward plot is indicative of nonspecific binding, negative cooperativity, or multiple classes of binding sites.
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