Structural and enzymatic insights into species-specific resistance to schistosome parasite drug therapy.

The antischistosomal prodrug oxamniquine is activated by a sulfotransferase (SULT) in the parasitic flatworm Schistosoma mansoni. Of the three main human schistosome species, only S. mansoni is sensitive to oxamniquine therapy despite the presence of SULT orthologs in Schistosoma hematobium and Schistosoma japonicum The reason for this species-specific drug action has remained a mystery for decades. Here we present the crystal structures of S. hematobium and S. japonicum SULTs, including S. hematobium SULT in complex with oxamniquine. We also examined the activity of the three enzymes in vitro; surprisingly, all three are active toward oxamniquine, yet we observed differences in catalytic efficiency that implicate kinetics as the determinant for species-specific toxicity. These results provide guidance for designing oxamniquine derivatives to treat infection caused by all species of schistosome to combat emerging resistance to current therapy.

Structural and Functional Characterization of the Enantiomers of the Antischistosomal Drug Oxamniquine.

BACKGROUND: For over two decades, a racemic mixture of oxamniquine (OXA) was administered to patients infected by Schistosoma mansoni, but whether one or both enantiomers exert antischistosomal activity was unknown. Recently, a ~30 kDa S. mansoni sulfotransferase (SmSULT) was identified as the target of OXA action. METHODOLOGY/PRINCIPAL FINDINGS: Here, we separate the OXA enantiomers using chromatographic methods and assign their optical activities as dextrorotary [(+)-OXA] or levorotary [(-)-OXA]. Crystal structures of the parasite enzyme in complex with optically pure (+)-OXA and (-)-OXA) reveal their absolute configurations as S- and R-, respectively. When tested in vitro, S-OXA demonstrated the bulk of schistosomicidal activity, while R-OXA had antischistosomal effects when present at relatively high concentrations. Crystal structures R-OXA•SmSULT and S-OXA•SmSULT complexes reveal similarities in the modes of OXA binding, but only the S-OXA enantiomer is observed in the structure of the enzyme exposed to racemic OXA. CONCLUSIONS/SIGNIFICANCE: Together the data suggest the higher schistosomicidal activity of S-OXA is correlated with its ability to outcompete R-OXA binding the sulfotransferase active site. These findings have important implications for the design, syntheses, and dosing of new OXA-based antischistosomal compounds.

The comparative study of a laccase-natural clinoptilolite-based catalyst activity and free laccase activity on model compounds.

For the first time a laccase from Trametes versicolor was immobilized on a natural clinoptilolite with Si/Al=5 to obtain a biocatalyst for environmental applications. Immobilization procedures exploiting adsorption and covalent binding were both tested, and only the last provided enough activity for practical applications. The optimal conditions for the immobilization of the enzyme on the support and the kinetic parameters for the free and covalent bonded laccase were determined. The laccase bonded to the zeolitic support showed a lower activity than the free laccase, but the pH and thermal stability were greater. 20 mg of dry biocatalyst containing 1 U of laccase were able to remove in 50h 73-78% of 2-chlorophenol and 2,4-dichlorophenol in relatively concentrated aqueous solutions (100 µmol L(-1)).

A study of the interaction between humic acids and acidic metabolites of phenanthrene by capillary electrophoresis.

Humic acids (HA) are able to interact with a wide range of pollulants and can influence their solubility, transport and bioavailability. In order to study the interaction between polar aromatic hydrocarbons and these macromolecules, an affinity capillary electrophoretic method, the Hummel-Dreyer (HD) method in its modified version, has been used. Two acidic metabolites of phenanthrene: 1-hydroxy-2-naphthoic acid (1-HNA) and 3,4-dihydroxybenzoic acid (3,4-DBA) were studied. The analysis for the binding studies was carried out by injecting a solution of HA in 25 mM ammonium hydrogen carbonate buffer (pH 9) into an uncoated fused-silica capillary, filled with buffer solutions of 1-HNA or 3,4-DBA in varying and increasing amounts. The results obtained indicate that both compounds bind to HA, as had been confirmed by dialysis experiments and literature data. CE proved to be a useful technique for investigating the link between xenobiotics and environmental macromolecules.