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Also, there has been some validation for the use of a single molecular conformation, where the estimation of binding affinities is based on either physical or statistical measures [8, 16, 17].
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Binding affinities are based on structural complementarity between proteins and binding sites, with molecular binding sites modeled by bit-strings.
ARF6 DNA-binding affinity was based on the peak score from the ARF6 ChIP-Seq analysis with CSAR.
A hierarchy of binding affinities was used, based on the in vitro experiments (Brittle et al., 2010; Simon et al., 2010), with phosphorylated Ft (FtP) and unphosphorylated Ds (Ds) producing the strongest partnership, and unphosphorylated Ft (Ft) and phosphorylated Ds (DsP) producing the weakest partnership (see 'Materials and methods').
Despite its simplicity, this type of selection model based on biophysical binding affinities is nontrivial from a population-genetic viewpoint since it leads to generic correlations between frequencies of nucleotides a i and a j within a site, see the Results section below.
The distance cutoff in the scoring function defining which pairwise interactions are taken into account when estimating the binding affinity was estimated based on a benchmark set of MHC class I binding data described in the methods section.
An alternate formulation based upon brain AR and gonad AR with different binding affinities was tried first.
Region-specific binding affinities were analyzed using Student's unpaired t-test.
Linear correlations between the predicted binding scores and the experimentally measured binding affinities were observed.
Thus native binding affinities are measured.
The highest binding affinities are observed with full exposure of the aptamer sequence in the loop, while duplex formation reduces binding affinity most likely due to the thermodynamics of DNA base pairing.
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