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Characterization of Carbonic Anhydrase Thermal Stability

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Carbonic Anhydrase as Drug Target

Abstract

Proteins have different stabilities in various media. The twelve carbonic anhydrase (CA) isoforms have been recombinantly prepared and used in the search of high-affinity ligands. It was important to characterize their thermal stability and determine solution conditions where the biochemical inhibition and biophysical interaction experiments could be optimally performed. Optimal conditions for the thermal stability experiments were determined and used in the search of inhibitors as described in further chapters. The stability of the CAs could be enhanced by the presence of inhibitors depending on the affinity and compound concentration. The main method used to determine the affinities of compounds in this book was the thermal shift assay, based on the increased stability of the protein by the binding ligand as described in Chap. 5. Here we describe the stabilities of all 12 CA isoforms in various buffers, salts, pH, osmolytes, organic solvents, and other excipients that are not specific binders and inhibitors of CAs.

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Authors and Affiliations

  1. Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Life Sciences Center, Vilnius University, Vilnius, Lithuania

    Asta Zubrienė & Daumantas Matulis

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  1. Asta Zubrienė
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  2. Daumantas Matulis
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Correspondence to Daumantas Matulis .

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Editors and Affiliations

  1. Department of Biothermodynamics and Drug Design, Vilnius University, Life Sciences Center, Institute of Biotechnology, Vilnius, Lithuania

    Daumantas Matulis

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Zubrienė, A., Matulis, D. (2019). Characterization of Carbonic Anhydrase Thermal Stability. In: Matulis, D. (eds) Carbonic Anhydrase as Drug Target. Springer, Cham. https://doi.org/10.1007/978-3-030-12780-0_4

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