Abstract
In this chapter we describe the model that quantifies interactions between proteins and ligands applying the thermal shift assay. When combined with fluorescence-based measurements of protein thermal unfolding, this model forms the basis of the fluorescent thermal shift assay—a widely applicable and cost-effective technique to quantify ligand affinity towards proteins. Most ligands stabilize proteins against thermal denaturation and shift their melting points towards higher temperatures. Equations that relate the shift in protein melting temperature with the ligand concentration are presented. The assay has been used to determine affinities of various sulfonamide inhibitor binding to carbonic anhydrase isoforms. The results illustrate applicability and limitations of the fluorescent thermal shift assay.
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Petrauskas, V., Zubrienė, A., Todd, M.J., Matulis, D. (2019). Inhibitor Binding to Carbonic Anhydrases by Fluorescent Thermal Shift Assay. In: Matulis, D. (eds) Carbonic Anhydrase as Drug Target. Springer, Cham. https://doi.org/10.1007/978-3-030-12780-0_5
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DOI: https://doi.org/10.1007/978-3-030-12780-0_5
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