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Despite the significantly weaker binding of p73CT compared to ΔNp73β to DNA, the DNA-binding specificity of both p73 proteins was identical.
It is possible that alloxan degraded to alloxanic acid over the course of the experiment, but it should also be noted that the two substrate analogues reported here showed significantly weaker binding to XcOGT than their observed inhibition of hOGT.
The IC50 of DD2 for the R82Q mutant was roughly 3 4-fold higher than that for the wild-type enzyme, suggesting a significantly weaker binding to the DD2 pocket.
However the effective Kd,individual from F1374 within Nup153AGPxAG,F1374 reveals significantly weaker binding for this interaction site than for F1374 when situated within the wild-type (WT) protein (Kd,individual = 7.3 mM compared to 0.8 mM, Figure 3).
Such a "scrambled" 6-nt oligonucleotide ssDNA sequence resulted in significantly weaker binding to N- HvGR-RBP1 (∼550 μM Kd compared to equilibrium dissociation constants of ∼4 43 μM for other ssDNA ligands), indicating that the nucleotides adjacent to the CTG element are indeed important modulators of the RRM binding affinity for its ssDNA target.
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Surface plasmon resonance and mass spectral studies shows the formation of an initial strong 1 1 complex followed by a significantly weaker secondary binding.
This revealed cross-reactivity with the K subfamily TIR, which was significantly weaker than binding to the cognate TIR.
The binding of chelerythrine to the ds poly(A) was significantly weaker than its binding to ss poly(A).
Using a biotin-avidin affinity assay to confirm the in vivo interaction between L22 and the RNAs used in our subsequent in vitro binding assays, we observed strong binding of L22 to EBER-1 and binding, albeit significantly weaker, to 28S rRNA (Fig. 1).
We observed specific NDE1 NDEL1 interaction and NDEL1 self-association, although their binding was significantly weaker than the noted DISC1 NDEL1 interaction (Fig. 5A).
The data are consistent with the affinity of dT27 being not much lower than that of dT35, but dT20 binding is significantly weaker.
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