NMR studies of p7 protein from hepatitis C virus.
The p7 protein of hepatitis C virus (HCV) plays an important role in the viral lifecycle. Like other members of the viroporin family of small membrane proteins, the amino acid sequence of p7 is largely conserved over the entire range of genotypes, and it forms ion channels that can be blocked by a number of established channel-blocking compounds. Its characteristics as a membrane protein make it difficult to study by most structural techniques, since it requires the presence of lipids to fold and function properly. Purified p7 can be incorporated into phospholipid bilayers and micelles. Initial solid-state nuclear magnetic resonance (NMR) studies of p7 in 14-O-PC/6-O-PC bicelles indicate that the protein contains helical segments that are tilted approximately 10 degrees and 25 degrees relative to the bilayer normal. A truncated construct corresponding to the second transmembrane domain of p7 is shown to have properties similar to those of the full-length protein, and was used to determine that the helix segment tilted at 10 degrees is in the C-terminal portion of the protein. The addition of the channel blocker amantadine to the full-length protein resulted in selective chemical shift changes, demonstrating that NMR has a potential role in the development of drugs targeted to p7.
Cook
Gabriel A
GA
Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA 92093, USA.
Opella
Stanley J
SJ
D000547:Amantadine / Q000737:chemistry; D000998:Antiviral Agents / Q000737:chemistry; D004926:Escherichia coli; D016174:Hepacivirus / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry; D008823:Micelles; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D010742:Phospholipid Ethers / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry; D017433:Protein Structure, Secondary; D011522:Protons; D014764:Viral Proteins / Q000737:chemistry*; D014867:Water / Q000737:chemistry; D000086082:Viroporin Proteins
D000547:Amantadine; D000998:Antiviral Agents; D008051:Lipid Bilayers; D008823:Micelles; D009587:Nitrogen Isotopes; D010742:Phospholipid Ethers; D010743:Phospholipids; D011522:Protons; D014764:Viral Proteins; D014867:Water; C052298:1,2-dihexadecyl-sn-glycero-3-phosphocholine; C088369:p7 protein, Hepatitis C virus; D000086082:Viroporin Proteins