0000000000352747
AUTHOR
Maria-josé Martínez-ferrer
Thermodynamic parameters of the interaction between Co(II) bovine carbonic anhydrase and anionic inhibitors
The pH dependence of the apparent affinity constants of perchlorate for cobalt(II)bovine carbonic anhydrase II has been measured by electronic absorption spectroscopy. The obtained data have been analyzed in terms of the ionization of two acidic groups of CoBCAII, and the affinity of perchlorate for the two water-containing species of the enzyme have been estimated. Furthermore, the affinity constants of nitrate, perchlorate, and azide for CoBCAII in the temperature range 5 degrees C-30 degrees C have been determined by spectrophotometric titrations at pH 7. The affinity constants for these ligands decrease with increasing temperatures. The temperature dependence of binding was used to esti…
1H 2D-NMR characterization of Ni(II)-substituted azurin fromPseudomonas aeruginosa
1 H two-dimemional NMR experiments on nickel(II)-substituted azurin have been succesfully applied. Despite the short relaxation time of the hyperfine-shifted resonances, the combined use of NOESY and COSY spectra allowed the alignment of 15 resonances belonging to the metal-coordinated residues Gly-45, His-46, His-117 and Met-121. Even in the case of the two broad and furthest downfield resonances, the NOESY spectra were successful in assigning these signals to the β-CH 2 protons of Cys-112. The protons of the non-coordinated residues Met-13, Phe-15 and Trp-48 were also assigned via NOESY, COSY and TOCSY experiments
1H NMR and UV-vis spectroscopic characterization of sulfonamide complexes of nickek(II)-carbonic anhydrase. Resonance assignments based on NOE effects
The binding of acetazolamide, p-fluorobenzensulfonamide, p-toluenesulfonamide, and sulfanilamide to nickel(II)-substituted carbonic anhydrase II has been studied by 1H NMR and electronic absorption spectroscopies. These inhibitors bind to the metal ion forming 1:1 complexes and their affinity constants were determined. The 1H NMR spectra of the formed complexes show a number of isotropically shifted signals corresponding to the histidine ligands. The complexes with benzene-sulfonamides gave rise to very similar 1H NMR spectra. The NMR data suggest that these aromatic sulfonamides bind to the metal ion altering its coordination sphere. In addition, from the temperature dependence of 1H NMR s…