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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Acknowledgments
The authors thank members of the Fuentes laboratory for helpful comments. E.J.F. is supported by National Institutes of Health grant R21-AI135305.
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Sun, Y.J., Fuentes, E.J. (2021). A Fluorescence-Based Assay to Determine PDZ–Ligand Binding Thermodynamics. In: Borg, JP. (eds) PDZ Mediated Interactions. Methods in Molecular Biology, vol 2256. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1166-1_8
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DOI: https://doi.org/10.1007/978-1-0716-1166-1_8
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