A multi-step strategy to obtain crystals of the dengue virus RNA-dependent RNA polymerase that diffract to high resolution.
Dengue virus, a member of the Flaviviridae genus, causes dengue fever, an important emerging disease with several million infections occurring annually for which no effective therapy exists. The viral RNA-dependent RNA polymerase NS5 plays an important role in virus replication and represents an interesting target for the development of specific antiviral compounds. Crystals that diffract to 1.85 A resolution that are suitable for three-dimensional structure determination and thus for a structure-based drug-design program have been obtained using a strategy that included expression screening of naturally occurring serotype variants of the protein, the addition of divalent metal ions and crystal dehydration.
Yap
Thai Leong
TL
Novartis Institute for Tropical Diseases, 10 Biopolis Road, Chromos Building, Singapore 138670, Singapore.
Chen
Yen Liang
YL
Xu
Ting
T
Wen
Daying
D
Vasudevan
Subhash G
SG
Lescar
Julien
J
D002384:Catalysis; D002413:Cations, Divalent; D002712:Chlorides / Q000737:chemistry; D002852:Chromatography, Ion Exchange; D003460:Crystallization; D018360:Crystallography, X-Ray; D003716:Dengue Virus / Q000201:enzymology*; D015636:Magnesium Chloride / Q000737:chemistry; D017895:Manganese Compounds / Q000737:chemistry; D017434:Protein Structure, Tertiary; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry* / Q000302:isolation & purification; D014764:Viral Proteins / Q000737:chemistry*
D002413:Cations, Divalent; D002712:Chlorides; D017895:Manganese Compounds; D014764:Viral Proteins; D015636:Magnesium Chloride; D012324:RNA-Dependent RNA Polymerase; C025340:manganese chloride