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In contrast, the D246A variant shows 70-fold greater binding affinity for the CCCGGG substrate.
Isothermal titration calorimetry (ITC) studies reveal that MRSA DHDPR has ∼20-fold greater binding affinity for NADPH (Kd = 1.5 μM) relative to NADH (Kd = 29 μM).
We then used the reconstructed LBD structure to design new mono-, bi- and tricyclic amine-bearing compounds that might be expected to show greater binding affinity for the LBD.
For example, RhoC has a greater binding affinity for ROCK than RhoA [ 16].
LPAR3 has a greater binding affinity for 2-acyl-LPA with unsaturated fatty acids, whereas LPAR1 and LPAR2 are more responsive to saturated acyl chains [ 63].
Rad52 has greater binding affinity for ssDNA than dsDNA (46, 47) and appears to have two separate binding sites for each in the N-terminus of Rad52 (48).
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Experimental data have previously demonstrated Y98H to exhibit higher stability and greater binding affinities for HIF1α compared with Y98N (63).
We anticipate the generation of mutants with more than 100 greater binding affinities for BtR than the wild-type toxin by integrating multiple mutations into one molecule.
These correspond to an ∼170 fold greater binding affinity of PPi for the mutant enzyme, and an ∼320 fold decrease in the rate of pyrophosphorolysis for this enzyme.
The greater binding affinity of AIRE PHD1 for H3K4me0 peptides was confirmed by both tryptophan fluorescence spectroscopy and isothermal titration calorimetry (ITC), yielding dissociation constants of ∼4 μM, ∼20 μM and >0.5 mM for H3K4me0, H3K4me1 and H3K4me2, respectively (supplementary Fig S2C online; Table 1).
Natively lipidated ApoE3 has been shown to have a greater binding affinity to Aβ than ApoE4 [31].
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