title: Preparation and handling of hepatitis C viral proteins NS3 and NS5B for structural studies.
abstract: HCV is a small positive-strand RNA virus responsible for a considerable proportion of acute and chronic hepatitis in humans. Although all HCV enzymes are, in theory, equally appropriate for therapeutic intervention, the NS3-NS4A serine protease and the NS5B RNA-dependent RNA polymerase are the most popular targets from a drug-discovery perspective. A number of active-site inhibitors of the NS3 protease as well as allosteric inhibitors of the NS5B polymerase are being developed. We determined the crystal structures of complexes of NS3/NS4A/active-site inhibitor as well as NS5B/allosteric inhibitor to permit structure-based drug design and the efficient optimization of leads. The methods for obtaining such structures by crystal soaking procedures are described.
authors.affiliation: Istituto di Ricerche di Biologia Molecolare "P. Angeletti", Pomezia (Rome), Italy.
authors.forename: Stefania Di
authors.initials: SD
authors.lastname: Marco
authors.forename: Andrea
authors.initials: A
authors.lastname: Carfí
mesh_terms: D000595:Amino Acid Sequence; D053000:Analytic Sample Preparation Methods; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D016174:Hepacivirus; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; D012324:RNA-Dependent RNA Polymerase
chemical_list: D010455:Peptides; D011480:Protease Inhibitors; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase