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The results suggest that the binding is a cooperative event in which both the sulfonamide residue and the helix-loop-helix motif contribute to the overall affinity.
It is also possible that the substrate binding is a cooperative event that involves more than one Rrp4 subunit.
Since the experimental evidence indicates that both sites show similar reduction in affinity and that site II is occupied first (and that the binding is a cooperative process), we wished to investigate the mechanism leading to the lower affinity of site II.
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Hill analysis of the data for 3Q suggests that heparin binding is cooperative with an estimated KD of 34 μ m and a stoichiometry of approximately one heparin molecule per four Aβ1-40 molecules at saturation (i.e., 4.5 saccharide units per Aβ1-40 monomer).
Since the screened compounds are A2AR antagonists, competition curves were expected to be monophasic, assuming that antagonist binding is not cooperative.
We assumed that HSP47 binding is not cooperative and that native procollagen has n independent binding sites i = 1,2, while unfolded procollagen has m binding sites j = 1,2.ach binding site i has the dissociation constant Kif and each binding site j has the dissociation constant Kiu.
It was found that the Ca2+ concentration for half-maximal binding activity of Dsg 1 is 0.8 mM Ca2+ and that binding is highly cooperative with the Hill coefficient being ≥5 (Waschke et al. 2007).
We cannot tell from our data whether PIP2 binding is cooperative (as the Hill model assumes) or non-cooperative (e.g., if multiple PIP2 molecules must bind, but independently, to open the channel).
If binding is cooperative, n = 2 may be more appropriate.
Therefore, we are assuming that 3A binding is cooperative, and that incomplete complexes have much higher affinity for 3A than does the reaction to form a new complex.
We have clarified that the affinity measured represents the sum of two distinct affinities, of which CT domain binding is cooperative.
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