Intrinsic thermodynamics of high affinity inhibitor binding to recombinant human carbonic anhydrase IV
| Autor: | David D. Timm, Jelena Jachno, Saulius Gražulis, Vaida Linkuvienė, Lina Baranauskienė, Asta Zubrienė, Alexey Smirnov, Jurgita Revuckienė, Abdul Waheed, Vilma Michailovienė, Aurelija Mickevičiūtė, Enrico Di Cera, Edita Čapkauskaitė, William S. Sly, Zhiwei Chen, Elena Manakova, Daumantas Matulis, Marius Gedgaudas, Jurgita Matulienė |
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| Rok vydání: | 2017 |
| Předmět: |
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Molecular 0301 basic medicine Thermal shift assay Stereochemistry Inorganic chemistry Biophysics Protonation Ligands Article 03 medical and health sciences chemistry.chemical_compound Carbonic Anhydrase IV Deprotonation Catalytic Domain Carbonic anhydrase Humans Carbonic Anhydrase Inhibitors biology Active site Isothermal titration calorimetry General Medicine Ligand (biochemistry) Recombinant Proteins 030104 developmental biology chemistry biology.protein Thermodynamics Hydroxide Protein Binding |
| Zdroj: | European Biophysics Journal. 47:271-290 |
| ISSN: | 1432-1017 0175-7571 |
| DOI: | 10.1007/s00249-017-1256-0 |
| Popis: | Membrane-associated carbonic anhydrase (CA) isoform IV participates in carbon metabolism and pH homeostasis and is implicated in the development of eye diseases such as retinitis pigmentosa and glaucoma. A series of substituted benzenesulfonamides were designed and their binding affinity to CA IV was determined by fluorescent thermal shift assay and isothermal titration calorimetry (ITC). Compound [(4-chloro-2-phenylsulfanyl-5-sulfamoyl-benzoyl)amino]propyl acetate (19) bound CA IV with the K d of 1.0 nM and exhibited significant selectivity over the remaining 11 human CA isoforms. The compound could be developed as a drug targeting CA IV. Various forms of recombinant CA IV were produced in Escherichia coli and mammalian cell cultures. Comparison of their temperature stability in various buffers and salt solutions demonstrated that CA IV is most stable at slightly alkaline conditions and at elevated sodium sulfate concentrations. High-resolution X-ray crystallographic structures of ortho-Cl and meta-thiazole-substituted benzene sulfonamide in complex with CA IV revealed the position of and interactions between the ligand and the protein. Sulfonamide inhibitor binding to CA IV is linked to several reactions—the deprotonation of the sulfonamide amino group, the protonation of CA–Zn(II)-bound hydroxide at the active site of CA IV, and the compensating reactions of the buffer. The dissection of binding-linked reactions yielded the intrinsic thermodynamic parameters, characterizing the interaction between CA IV and the sulfonamides in the binding-able protonation forms, including Gibbs energy, enthalpy, and entropy, that could be used for the characterization of binding to any CA in the process of drug design. |
| Databáze: | OpenAIRE |
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