Crystal structure of a novel dimeric form of NS5A domain I protein from hepatitis C virus
- PMID: 19244328
- PMCID: PMC2668466
- DOI: 10.1128/JVI.02352-08
Crystal structure of a novel dimeric form of NS5A domain I protein from hepatitis C virus
Abstract
A new protein expression vector design utilizing an N-terminal six-histidine tag and tobacco etch virus protease cleavage site upstream of the hepatitis C virus NS5A sequence has resulted in a more straightforward purification method and improved yields of purified NS5A domain I protein. High-resolution diffracting crystals of NS5A domain I (amino acids 33 to 202) [NS5A(33-202)] were obtained by using detergent additive crystallization screens, leading to the structure of a homodimer which is organized differently from that published previously (T. L. Tellinghuisen, J. Marcotrigiano, and C. M. Rice, Nature 435:374-379, 2005) yet is consistent with a membrane association model for NS5A. The monomer-monomer interface of NS5A(33-202) features an extensive buried surface area involving the most-highly conserved face of each monomer. The two alternate structural forms of domain I now available may be indicative of the multiple roles emerging for NS5A in viral RNA replication and viral particle assembly.
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References
-
- Blight, K. J., A. A. Kolykhalov, and C. M. Rice. 2000. Efficient initiation of HCV RNA replication in cell culture. Science 2901972-1974. - PubMed
-
- Brass, V., E. Bieck, R. Montserret, B. Wolk, J. A. Hellings, H. E. Blum, F. Penin, and D. Moradpour. 2002. An amino-terminal amphipathic alpha-helix mediates membrane association of the hepatitis C virus nonstructural protein 5A. J. Biol. Chem. 2778130-8139. - PubMed
-
- Brodsky, O., and C. N. Cronin. 2006. Economical parallel protein expression screening and scale-up in Escherichia coli. J. Struct. Funct. Genomics 7101-108. - PubMed
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