Inhibitor binding to metal-substituted metalloenzyme: Sulfonamide affinity for carbonic anhydrase IX.

Transition metal ions are structural and catalytic cofactors of many proteins including human carbonic anhydrase (CA), a Zn-dependent hydrolase. Sulfonamide inhibitors of CA recognize and form a coordination bond with the Zn ion located in the active site of the enzyme. The Zn ion may be removed or substituted with other metal ions. Such CA protein retains the structure and could serve as a tool to study metal ion role in the recognition and binding affinity of inhibitor molecules. We measured the affinities of selected divalent transition metal ions, including Mn, Fe, Co, Ni, Cu, Cd, Hg, and Zn to metal-free CA isozymes CA I, CA II, and CAIX by fluorescence-based thermal shift assay, prepared metal-substituted CAs, and determined binding of diverse sulfonamide compounds. Sulfonamide inhibitor binding to metal substituted CA followed a U-shape pH dependence. The binding was dissected to contributing binding-linked reactions and the intrinsic binding reaction affinity was calculated. This value is independent of pH and protonation reactions that occur simultaneously upon binding native CA and as demonstrated here, to metal substituted CA. Sulfonamide inhibitor binding to cancer-associated isozyme CAIX diminished in the order: Zn > Co > Hg > Cu > Cd > Mn > Ni. Energetic contribution of the inhibitor-metal coordination bond was determined for all above metals. The understanding of the principles of metal influence on ligand affinity and selectivity should help design new drugs targeting metalloenzymes.

Relation between thickness, crystallite size and magnetoresistance of nanostructured La1- x Sr x Mn y O3±Î´ films for magnetic field sensors.

In the present study the advantageous pulsed-injection metal organic chemical vapour deposition (PI-MOCVD) technique was used for the growth of nanostructured La1- x Sr x Mn y O3±Î´ (LSMO) films on ceramic Al2O3 substrates. The compositional, structural and magnetoresistive properties of the nanostructured manganite were changed by variation of the processing conditions: precursor solution concentration, supply frequency and number of supply sources during the PI-MOCVD growth process. The results showed that the thick (≈400 nm) nanostructured LSMO films, grown using an additional supply source of precursor solution in an exponentially decreasing manner, exhibit the highest magnetoresistance and the lowest magnetoresistance anisotropy. The possibility to use these films for the development of magnetic field sensors operating at room temperature is discussed.