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The holdup assay was carried out against the biotinylated peptide PBM-HBc (peptide sequence -RRRRSQSRESQC) in duplicates as previously described48, 49 with minor modifications. We measured PBM-HBc interactions against 255 human PDZ domains. The minimal binding intensity (BI) threshold value is 0.2 to define a significant interaction, as previously reported48.
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