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  • Title: A Fluorescence-Based Assay to Determine PDZ-Ligand Binding Thermodynamics.
    Author: Sun YJ, Fuentes EJ.
    Journal: Methods Mol Biol; 2021; 2256():137-148. PubMed ID: 34014520.
    Abstract:
    Postsynaptic density-95, disks-large, and zonula occludens-1 (PDZ) domain interactions with cognate linear binding motifs (i.e., PDZ-binding motifs or PBMs) are important for many biological processes and can be pathological when disrupted. There are hundreds of PDZ-PBM interactions reported but few have been quantitatively determined. Moreover, PDZ-PBM interactions have been identified as potential therapeutic targets. To thoroughly understand PDZ-PBM binding energetics and their specificity, we have developed a sensitive and quantitative equilibrium binding assay. Here, we describe a protocol for determining PDZ-PBM binding energetics using fluorescence anisotropy-based methodology.
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