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The apicoplast and the proteins present therein are parasite-specific targets for chemotherapy of apicomplexan parasites. Ferredoxin-NADP(+) reductase (FNR) is an important enzyme present in the apicoplast of Toxoplasma gondii that operates as a general electron switch at the bifurcation step of many different electron transfer pathways. In spite of its importance as drug target not much structural information on the enzyme is available. Using fluorescence and CD spectroscopy in combination with enzyme activity measurement and size exclusion chromatography, we studied the pH-dependent changes in structural and functional properties and interdomain interactions in recombinant Toxoplasma gondii ferredoxin-NADP(+) reductase (TgFNR) to understand the interactions responsible for stabilization of native conformation and modulation of functional activity of the enzyme. Under physiological conditions, the recombinant TgFNR is stabilized in an open conformation. The open conformation of the enzyme was found to be essential for its optimum functioning, as induction of compactness/rigidity by modulation of pH, leads to decrease in the functional activity. In native conformation, strong interactions exist between the NADP(+)- and FAD-binding domains thus making the enzyme a structurally cooperative molecule. Under acidic conditions (pH about 4), the interdomain interactions present in native TgFNR were lost and the enzyme became structurally noncooperative. The pH-induced structural alterations in the NADP(+) binding domain, more precisely compaction of the conformation lead to its stabilization against thermal denaturation. The studies demonstrate the significance of electrostatic interactions both in stabilization of native conformation and maintenance of structural cooperativity in TgFNR.

Background: α-isopropylmalate synthase (MtαIPMS), an enzyme that catalyzes the first committed step of the leucine biosynthetic pathway of Mycobacterium tuberculosis is a potential drug target for the anti-tuberculosis drugs. Cations induce differential effect of activation and inhibition of MtαIPMS. To date no concrete mechanism for such an opposite effect of similarly charged cations on the functional activity of enzyme has been presented.

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Submerged arc welding of SA 516 grade 60 pressure vessel grade steel was conducted with different heat plate thicknesses and the influence of cooling rate on microstructure, Vickers hardness, and impact toughness of heat affected zone (HAZ) of weldment was systematically investigated. Weld cooling rates vary with change in heat input or variation in plate thickness of base metal. Results showed that thin plates accumulate the heat, which cause grain coarsening and loss of acicular ferrite (AF) microstructure, which is further responsible for lower impact strength of welded joint. It is deemed that faster cooling rates due to heat sink in thickness direction with thick plates cause high percentage of AF with finer grain and enhanced hardness values. Improved impact strength with thick plates with same heat input signifies that supplying heat more than required to thin plates may cause microstructural deterioration and responsible for impact strength loss of weldments. Test demonstrates that the cooling rate should be above 15 °C/s to keep impact strength loss within considerable limits.

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