These structural modifications affect the affinity of the TTR-binding site not only to T4 but also to exogenous compounds, and explain the differences in the TTR-binding potency of compounds reported between species.
Overall, we conclude that the FP assay was appropriate and efficient to evaluate the binding potency of the tested compounds and dust extracts.
Briefly, we used a commercially available high-throughput ligand binding assay (PolarScreen™ PPARγ-Competitor Assay Kit; Invitrogen, Carlsbad, CA) to investigate the binding potency of the tested compounds to PPARγ LBD.
In this approach, an affinity fingerprint is the pattern of the in vitro binding potency of a single compound to a reference panel of eight diverse proteins.
Using a database of such affinity fingerprints, the authors were able to predict the binding potency of a novel compound for a specific protein target using a multivariate linear regression model, derived from the affinity fingerprints of a small set of training compounds.
It might be possible that compounds in dust, such as TBB, TBPH, PBDEs, and DEHP, could be metabolized to PPARγ-active ligands after incubation, which was supported by the increased binding potency of the prepared MIX containing these chemicals.
Although some of the tested compounds (e.g., TBEP and TBuP) showed weaker PPARγ binding potency, these compounds may yet be of great concern because of their ubiquitous detection in indoor environments, with levels up to micrograms to milligrams per gram of dust (Van den Eede et al. 2011).
This study also recommended using disrupted circulation and transport of TH via the binding to serum transport proteins for the evaluation of the TH-disrupting potency of compounds.
In most cases, the potency of compounds was consistent with the picture of ligand binding derived from the reported X-ray structures.
A dose-response experiment was performed to determine the potency of compound 1 at inhibiting the DNA-binding activity of STAT3.
