Articles by Marilyne Blemont in JoVE
Hochdurchsatz-Screening Quantitative Expression und Aufreinigung rekombinanter zu Disulfid-reichen Proteine Venom Produziert in Applied Natalie J. Saez1, Hervé Nozach2, Marilyne Blemont1, Renaud Vincentelli1 1Architecture et Fonction des Macromolécules Biologiques (AFMB), Aix-Marseille Université, 2iBiTec-S, Service d'Ingénierie Moléculaire des Protéines (SIMOPRO), Commissariat à l'énergie atomique et aux énergies alternatives (CEA) Saclay, France Ein Protokoll für die quantitative, Expressionsscreening mit hohem Durchsatz und analytischen Reinigung von Fusionsproteinen aus kleinen Escherichia coli-Kulturen beschrieben und auf die Expression von Disulfid-reichen Tiergift-Protein-Targets aufgebracht.
Other articles by Marilyne Blemont on PubMed
Identification of Allosteric Inhibitors Blocking the Hepatitis C Virus Polymerase NS5B in the RNA Synthesis Initiation Step Antiviral Research. Oct, 2009 | Pubmed ID: 19589358 Hepatitis C virus (HCV) RNA-dependent RNA polymerase NS5B constitutes a target of choice for the development of anti-HCV drugs. Although many small molecules have been identified as allosteric inhibitors of NS5B, very few are active in clinical applications. We have screened 17,000 compounds in an enzymatic assay involving the purified NS5B in order to increase the therapeutic arsenal. We hoped to shed some light on the precise mechanism of RNA synthesis. We succeeded in isolating a series of 21 original inhibitors of the RNA synthesis by NS5B. Four of these non-nucleoside inhibitors (NNIs) could be mapped to the known binding site called 'B' as judged by the decrease in their inhibition potency when assayed with a 'B' site mutant, M423T NS5B. Incidentally, our in silico model pointed to Y477 as a key residue for inhibitor binding. In vitro, Y477F mutant loses its sensitivity to the newly discovered inhibitors but is unable to extend primers during the elongation phase. Our results demonstrate that elements of the 'B' site are involved in the conformational changes required in the switch between the different RNA synthesis steps and that compounds targeting this site could lock the enzyme in its initiation phase.