Backbone H(N), N, C(alpha), C', and C(beta) assignment of the 6-phosphogluconolactonase, a 266-residue enzyme of the pentose-phosphate pathway from human parasite Trypanosoma brucei

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Effet de la fonctionnalisation d'aciers inoxydables sur l'adhésion de films organiques

International audienceDans l'industrie automobile, le traitement par phosphatation est utilisé comme primaire d'adhérence avant application d’une peinture cataphorétique. La phosphatation engendre une grande production d'effluents, de boues métalliques, qui nécessitent une station de traitement. De plus, la mise en oeuvre du bain est relativement contraignante car plusieurs rinçages en cascade sont nécessaires. Par ailleurs, le bain est instable dans le temps, ce qui nécessite des analyses quotidiennes et donc la présence d'un technicien. Concernant la toxicité du bain, il est relativement dangereux par la présence d'acides concentrés, de nitrates et de nitrites. Pour finir, nous sommes dans une ère de miniaturisation, on peut donc trouver des problèmes d'assemblage car ce revêtement est de l'ordre du micromètre (2-30 μm)

Biochemical characterization and NMR studies of the nucleotide-binding domain 1 of multidrug-resistance-associated protein 1: evidence for interaction between ATP and Trp653

International audienceMultidrug-resistance-associated protein 1 (MRP1/ABCC1) is a human ATP-binding cassette transporter that confers cell resistance to antitumour drugs. Its NBDs (nucleotide-binding domains) bind/hydrolyse ATP, a key step in the activation of MRP1 function. To relate its intrinsic functional features to the mechanism of action of the full-size transporter, we expressed the N-terminal NBD1 domain (Asn(642) to Ser(871)) in Escherichia coli. NBD1 was highly purified under native conditions and was characterized as a soluble monomer. (15)N-labelling allowed recording of the first two-dimensional NMR spectra of this domain. The NMR study showed that NBD1 was folded, and that Trp(653) was a key residue in the NBD1-ATP interaction. Thus, interaction of NBD1 with ATP/ADP was studied by intrinsic tryptophan fluorescence. The affinity for ATP and ADP were in the same range (K (d(ATP))=118 microM and K (d(ADP))=139 microM). Binding of nucleotides did not influence the monomeric state of NBD1. The ATPase activity of NBD1 was magnesium-dependent and very low [V (max) and K (m) values of 5x10(-5) pmol of ATP x (pmol NBD1)(-1) x s(-1) and 833 microM ATP respectively]. The present study suggests that NBD1 has a low contribution to the ATPase activity of full-length MRP1 and/or that this activity requires NBD1-NBD2 heterodimer formation

Crustal deformation at the southernmost part of the Ryukyu subduction (East Taiwan) as revealed by new marine seismic experiments

International audienceThe southernmost part of the Ryukyu subduction, where the Philippine Sea Plate is subducting under the Eurasian Plate, is known to be a very seismically active region of transition from a north-dipping subduction along the Ryukyu subduction to an ~ SE-NW collision along the Taiwanese orogenic wedge. In this paper, we will focus on the Ryukyu forearc area close to Taiwan where the deformation is paroxysmal. In order to decipher the nature of the seismic deformation in this region, a three month passive experiment, combining 22 Ocean Bottom Seismometers and 51 onland stations, has been led. Starting from an a-priori heterogeneous model, we have obtained 801 well-located earthquake hypocenters, a precise P-wave tomography model and 14 focal mechanisms. The seismicity along the Ryukyu forearc is mainly located not only in the vicinity of the Interplate Seismogenic Zone (ISZ) but also within both the subducting PSP and the overriding plate. Seismicity within the upper-plate is essentially localized east of Nanao basin where NW-SE extension occurs, and northwest of the Hoping basin where strike-slip dominates. As revealed by both the P-wave velocity structure and the newly derived seismicity, we argue that a sub-vertical step offsetting the subducting PSP around 10 km may support the presence of a trench-parallel tear. The PSP also undergoes extension in its upper part that is probably caused by buckling and slab pull. The P-wave velocity structure reveals three other major features: (1) a continuity between the Central Range and the Ryukyu Arc with a shallower Moho (~ 30 km depth) between ~ 122.3°N and ~ 122.5°N along the Ryukyu Arc, (2) high P-wave velocities along the eastern side of the Central Range and, (3) two bodies with similar high crustal velocities (6.5-7.0 km/s) at 12-18 km depths, embedded within the Ryukyu arc basement, just north of Hoping Basin and north of the Nanao Basin

The influence of self-assembled monolayers on the practical adhesion of a cataphoretic paint

International audienceThe objective of this study is to replace steel-phosphating treatments with self-assembled monolayers (SAMs), in this case, of phosphonic acids. Two industrial applications are targeted: obtaining an adhesion primer between steel and paint or lubricating stainless steel for mechanical applications, such as stamping ; research has been conducted using alkyl phosphonic acids. The carbon chain length and terminal function are changed to obtain the best properties depending on the application

Targeting of eEF1A With Amaryllidaceae Isocarbostyrils as a Strategy to Combat Melanomas

Melanomas display poor response rates to adjuvant therapies because of their intrinsic resistance to proapoptotic stimuli. This study indicates that such resistance can be overcome, at least partly, through the targeting of eEF1A elongation factor with narciclasine, an Amaryllidaceae isocarbostyril controlling plant growth. Narciclasine displays IC50 growth inhibitory values between 30–100 nM in melanoma cell lines, irrespective of their levels of resistance to proapoptotic stimuli. Normal noncancerous cell lines are much less affected. At nontoxic doses, narciclasine also significantly improves (P=0.004) the survival of mice bearing metastatic apoptosis-resistant melanoma xenografts in their brain. The eEF1A targeting with narciclasine (50 nM) leads to 1) marked actin cytoskeleton disorganization, resulting in cytokinesis impairment, and 2) protein synthesis impairment (elongation and initiation steps), whereas apoptosis is induced at higher doses only (≥200 nM). In addition to molecular docking validation and identification of potential binding sites, we biochemically confirmed that narciclasine directly binds to human recombinant and yeast-purified eEF1A in a nanomolar range, but not to actin or elongation factor 2, and that 5 nM narciclasine is sufficient to impair eEF1A-related actin bundling activity. eEF1A is thus a potential target to combat melanomas regardless of their apoptosis-sensitivity, and this finding reconciles the pleiotropic cytostatic of narciclasine