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The first group consisted of mutations that increased the experimental rate of association through electrostatic optimization.
Conversely, we have shown that crowding has only a small effect on the rate of association of protein complexes.
Mahadeo, D., Kaplan, L., Chao, M. V. & Hempstead, B. L. High affinity nerve growth factor binding displays a faster rate of association than p140trk binding.
Importantly, specificity is achieved without slowing the rate of association, an important requirement for rapid inhibition of the colicin in the producing bacterial cell.
Besides quantification of the interactions, the rate of association (ka) and dissociation (kd), the KA and KD standard equilibrium constants and the binding free energy (ΔG°) were also calculated on the basic of SPR measurements.
To gain further quantitative insight, we next analyzed the effect of HIF-2α mutations on the kinetics of HIF-2α peptide binding to pVHL via biolayer interferometry (BLI 41, which measures the rate of association and dissociation of an analyte molecule (purified VBC complex) to and from an immobilized bait molecule (biotinylated HIF-2αOH peptide).
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Despite the low rate of associations observed in the buccal data, out of the 96 sex-associated CpGs in the buccal data, 16 CpGs were also associated with sex in KORA F4.
The empirical type I error rate of association-mapping approaches based on adjusted entry means (two-step approaches) is only slightly higher than that of approaches in which the phenotypic data analysis and the association analysis were performed in one step (one-step approaches) [ 20].
Thus, the decrease in the rate of associations detected with incremental linkage distance or recombination rates was assessed considering the extreme points from θ = 0 for which all associations detected are true positive associations (power) and from θ = 0.0002 for which most associations detected are type I errors.
Instead it was determined indirectly as the ratio of the rates of association and dissociation measured by means of the associated conformational change.
In our BLI assay we similarly found polyanionic heparin reduced binding affinities and rates of association (Supplementary Fig. 11a).
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