Structure and reactivity of Trypanosoma brucei pteridine reductase: inhibition by the archetypal antifolate methotrexate

The protozoan Trypanosoma brucei has a functional pteridine reductase (TbPTR1), an NADPH-dependent short-chain reductase that participates in the salvage of pterins, which are essential for parasite growth. PTR1 displays broad-spectrum activity with pterins and folates, provides a metabolic bypass for inhibition of the trypanosomatid dihydrofolate reductase and therefore compromises the use of antifolates for treatment of trypanosomiasis. Catalytic properties of recombinant TbPTR1 and inhibition by the archetypal antifolate methotrexate have been characterized and the crystal structure of the ternary complex with cofactor NADP(+) and the inhibitor determined at 2.2 Å resolution. This enzyme shares 50% amino acid sequence identity with Leishmania major PTR1 (LmPTR1) and comparisons show that the architecture of the cofactor binding site, and the catalytic centre are highly conserved, as are most interactions with the inhibitor. However, specific amino acid differences, in particular the placement of Trp221 at the side of the active site, and adjustment of the β6-α6 loop and α6 helix at one side of the substrate-binding cleft significantly reduce the size of the substrate binding site of TbPTR1 and alter the chemical properties compared with LmPTR1. A reactive Cys168, within the active site cleft, in conjunction with the C-terminus carboxyl group and His267 of a partner subunit forms a triad similar to the catalytic component of cysteine proteases. TbPTR1 therefore offers novel structural features to exploit in the search for inhibitors of therapeutic value against African trypanosomiasis

Effectiveness of recovered magnesium phosphates as fertilizers in neutral and slightly alkaline soils

Magnesium phosphates such as struvite (MgNH4PO4 · 6H2O) can be recovered from municipal, industrial, and agricultural wastewaters. However, limited information is available on the beneficial reuse of these recovered products; research has focused on low pH soils. Th is study determined whether recovered struvite and dittmarite (MgNH4PO4 · H2O) were effective P fertilizers in neutral to slightly alkaline soils. In addition to commercially available triple superphosphate (TSP) and certified organic rock phosphate (RP), recovered struvite, dittmarite, and a heterogeneous recovered phosphate were evaluated in a laboratory dissolution study and as fertilizers for spring wheat (Triticum aestivum L.) in a greenhouse study. Struvite and dittmarite were much more soluble than RP, but less soluble than TSP. Laboratory dissolution kinetics were fast, with most materials nearing equilibrium within 7 to 14 d. At a soil pH of 6.5, both dittmarite and struvite increased the average plant P concentration over the control. Struvite and dittmarite performance was similar to TSP. There were no significant differences in plant dry matter (DM) production or total P uptake at pH 6.5. In the limed soil (pH 7.6), many treatments had plant P concentrations significantly lower than the control, but most fertilizers increased DM production over the control; all fertilizers generally performed similarly to one another. These findings support previous work showing recovered Mg phosphates to be effective in acidic soils, and provide evidence that they are also effective in slightly alkaline soils. Recovered Mg phosphates could become a useful alternative for P fertilization in arid and semiarid environments