Preparation and handling of hepatitis C viral proteins NS3 and NS5B for structural studies.
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.
Marco
Stefania Di
SD
Istituto di Ricerche di Biologia Molecolare "P. Angeletti", Pomezia (Rome), Italy.
Carfí
Andrea
A
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
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