title: Intrinsic thermodynamics of 4-substituted-2,3,5,6-tetrafluorobenzenesulfonamide binding to carbonic anhydrases by isothermal titration calorimetry.
abstract: Para substituted tetrafluorobenzenesulfonamides bind to carbonic anhydrases (CAs) extremely tightly and exhibit some of the strongest known protein-small ligand interactions, reaching an intrinsic affinity of 2 pM as determined by displacement isothermal titration calorimetry (ITC). The enthalpy and entropy of binding to five CA isoforms were measured by ITC in two buffers of different protonation enthalpies. The pKa values of compound sulfonamide groups were measured potentiometrically and spectrophotometrically, and enthalpies of protonation were measured by ITC in order to evaluate the proton linkage contributions to the observed binding thermodynamics. Intrinsic means the affinity of a sulfonamide anion for the Zn bound water form of CAs. Fluorination of the benzene ring significantly enhanced the observed affinities as it increased the fraction of deprotonated ligand while having little impact on intrinsic affinities. Intrinsic enthalpy contributions to the binding affinity were dominant over entropy and were more exothermic for CA I than for other CA isoforms. Thermodynamic measurements together with the X-ray crystallographic structures of protein-ligand complexes enabled analysis of structure-activity relationships in this enzyme ligand system.
authors.affiliation: Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Vilnius University, Graičiūno 8, Vilnius LT-02241, Lithuania. Electronic address: astzu@ibt.lt.
authors.forename: Asta
authors.initials: A
authors.lastname: Zubrienė
authors.affiliation: Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Vilnius University, Graičiūno 8, Vilnius LT-02241, Lithuania.
authors.forename: Joana
authors.initials: J
authors.lastname: Smirnovienė
authors.affiliation: Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Vilnius University, Graičiūno 8, Vilnius LT-02241, Lithuania.
authors.forename: Alexey
authors.initials: A
authors.lastname: Smirnov
authors.affiliation: Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Vilnius University, Graičiūno 8, Vilnius LT-02241, Lithuania.
authors.forename: Vaida
authors.initials: V
authors.lastname: Morkūnaitė
authors.affiliation: Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Vilnius University, Graičiūno 8, Vilnius LT-02241, Lithuania.
authors.forename: Vilma
authors.initials: V
authors.lastname: Michailovienė
authors.affiliation: Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Vilnius University, Graičiūno 8, Vilnius LT-02241, Lithuania.
authors.forename: Jelena
authors.initials: J
authors.lastname: Jachno
authors.affiliation: Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Vilnius University, Graičiūno 8, Vilnius LT-02241, Lithuania.
authors.forename: Vaida
authors.initials: V
authors.lastname: Juozapaitienė
authors.affiliation: Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Vilnius University, Graičiūno 8, Vilnius LT-02241, Lithuania.
authors.forename: Povilas
authors.initials: P
authors.lastname: Norvaišas
authors.affiliation: Department of Protein-DNA Interactions, Institute of Biotechnology, Vilnius University, Graičiūno 8, Vilnius LT-02241, Lithuania.
authors.forename: Elena
authors.initials: E
authors.lastname: Manakova
authors.affiliation: Department of Protein-DNA Interactions, Institute of Biotechnology, Vilnius University, Graičiūno 8, Vilnius LT-02241, Lithuania.
authors.forename: Saulius
authors.initials: S
authors.lastname: Gražulis
authors.affiliation: Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Vilnius University, Graičiūno 8, Vilnius LT-02241, Lithuania.
authors.forename: Daumantas
authors.initials: D
authors.lastname: Matulis
mesh_terms: D002151:Calorimetry; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000378:metabolism*; D019277:Entropy; D054879:Halogenation; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D007536:Isomerism; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D011522:Protons; D013449:Sulfonamides / Q000737:chemistry* / Q000378:metabolism*; D014867:Water / Q000737:chemistry; D015032:Zinc / Q000737:chemistry; D000096926:Benzenesulfonamides
chemical_list: D002256:Carbonic Anhydrases; D007527:Isoenzymes; D011522:Protons; D013449:Sulfonamides; D014867:Water; D015032:Zinc; D000096926:Benzenesulfonamides
keywords: Carbonic anhydrase inhibitors; Intrinsic thermodynamics of binding; Isothermal titration calorimetry; Structure–thermodynamics relationships; X-ray crystallography