Diuretic drug binding to human glutathione transferase P1-1: potential role of Cys-101 revealed in the double mutant C47S/Y108V

The diuretic drug ethacrynic acid (EA), both an inhibitor and substrate of pi class glutathione S-transferase (GST P1-1), has been tested in clinical trials as an adjuvant in chemotherapy. We recently studied the role of the active site residue Tyr-108 in binding EA to the enzyme and found that the analysis was complicated by covalent binding of this drug to the highly reactive Cys-47. Previous attempts to eliminate this binding by chemical modification yielded ambiguous results and therefore we decided here to produce a double mutant C47S/Y108V by site directed mutagenesis and further expression in Escherichia coli and the interaction of EA and its GSH conjugate (EASG) examined by calorimetric studies and X-ray diffraction. Surprisingly, in the absence of Cys-47, Cys-101 (located at the dimer interface) becomes a target for modification by EA, albeit at a lower conjugation rate than Cys-47. The Cys-47 → Ser mutation in the double mutant enzyme induces a positive cooperativity between the two subunits when ligands with affinity to G-site bind to enzyme. However, this mutation does not seem to affect the thermodynamic properties of ligand binding to the electrophilic binding site (H-site) and the thermal or chemical stability of this double mutant does not significantly affect the unfolding mechanism in either the absence or presence of ligand. Crystal structures of apo and an EASG complex are essentially identical with a few exceptions in the H-site and in the water network at the dimer interface

Carbon nanotubes

International audienceCarbon nanotubes (CNT s) are remarkable objects that once looked set to revolutionize the technological landscape in the near future. Since the 1990s and for twenty years thereafter, it was repeatedly claimed that tomorrow's society would be shaped by nanotube applications, just as silicon-based technologies dominate society today. Space elevators tethered by the strongest of cables, hydrogen-powered vehicles, artificial muscles: these were just a few of the technological marvels that we were told would be made possible by the science of carbon nanotubes. Of course, this prediction is still some way from becoming reality; most often the possibilities and potential have been evaluated, but actual technological development is facing the unforgiving rule that drives the transfer of a new material or a new device to market: profitability. New materials, even more so for nanomaterials, no matter how wonderful they are, have to be cheap to produce, constant in quality, easy to handle, and nontoxic. Those are the conditions for an industry to accept a change in its production lines to make them nanocompatible. Consider the example of fullerenes – molecules closely related to nanotubes. The anticipation that surrounded these molecules, first reported in 1985, resulted in the bestowment of a Nobel Prize for their discovery in 1996. However, two decades later, very few fullerene applications have reached the market, suggesting that similarly enthusiastic predictions about nanotubes should be approached with caution, and so should it be with graphene, another member of the carbon nanoform family which joined the game in 2004, again acknowledged by a Nobel Prize in 2010. There is no denying, however, that the expectations surrounding carbon nanotubes are still high, because of specificities that make them special compared to fullerenes and graphene: their easiness of production, their dual molecule/nano-object nature, their unique aspect ratio, their robustness, the ability of their electronic structure to be given a gap, and their wide typology etc. Therefore, carbon nanotubes may provide the building blocks for further technological progress, enhancing our standard of living. In this chapter, we first describe the structures, syntheses, growth mechanisms, and properties of carbon nanotubes. Then we introduce nanotube-based materials, which comprise on the one hand those formed by reactions and associations of all-carbon nanotubes with foreign atoms, molecules and compounds, and on the other hand, composites, obtained by incorporating carbon nanotubes in various matrices. Finally, we will provide a list of applications currently on the market, while skipping the potentially endless and speculative list of possible applications