title: Six-helix bundle assembly and characterization of heptad repeat regions from the F protein of Newcastle disease virus.
abstract: Paramyxoviruses may adopt a similar fusion mechanism to other enveloped viruses, in which an anti-parallel six-helix bundle structure is formed post-fusion in the heptad repeat (HR) regions of the envelope fusion protein. In order to understand the fusion mechanism and identify fusion inhibitors of Newcastle disease virus (NDV), a member of the Paramyxoviridae family, we have developed an E. coli system that separately expresses the F protein HR1 and HR2 regions as GST fusion proteins. The purified cleaved HR1 and HR2 have subsequently been assembled into a stable six-helix bundle heterotrimer complex. Furthermore, both the GST fusion protein and the cleaved HR2 show virus-cell fusion inhibition activity (IC(50) of 1.07-2.93 microM). The solubility of the GST-HR2 fusion protein is much higher than that of the corresponding peptide. Hence this provides a plausible method for large-scale production of HR peptides as virus fusion inhibitors.
authors.affiliation: Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
authors.forename: Ming
authors.initials: M
authors.lastname: Yu
authors.affiliation: Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
authors.forename: Enxiu
authors.initials: E
authors.lastname: Wang
authors.affiliation: Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
authors.forename: Youfang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, People's Republic of China2.
authors.forename: Dianjun
authors.initials: D
authors.lastname: Cao
authors.affiliation: Dept of Virus Research, University of Military Supplies, Changchun 130062, People's Republic of China3.
authors.forename: Ningyi
authors.initials: N
authors.lastname: Jin
authors.affiliation: Laboratory of Immunobiology, Dana-Farber Cancer Institute and Dept of Medicine, Harvard Medical School, Boston, MA 02115, USA4.
authors.forename: Catherine W-H
authors.initials: CW
authors.lastname: Zhang
authors.affiliation: Laboratory of Structural Biology and MOE Laboratory of Protein Sciences, Tsinghua University, Beijing 100084, People's Republic of China5.
authors.forename: Mark
authors.initials: M
authors.lastname: Bartlam
authors.affiliation: Laboratory of Structural Biology and MOE Laboratory of Protein Sciences, Tsinghua University, Beijing 100084, People's Republic of China5.
authors.forename: Zihe
authors.initials: Z
authors.lastname: Rao
authors.affiliation: Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
authors.forename: Po
authors.initials: P
authors.lastname: Tien
authors.affiliation: Laboratory of Molecular Medicine, Children's Hospital, Dept of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 320 Longwood Avenue, Boston, MA 02115, and Dept of Molecular and Cellular Biology, Harvard University, 7 Divinity Avenue, Cambridge, MA 02138, USA6.|Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
authors.forename: George F
authors.initials: GF
authors.lastname: Gao
mesh_terms: D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D002681:China; D002942:Circular Dichroism; D006367:HeLa Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D008561:Membrane Fusion / Q000187:drug effects; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009522:Newcastle disease virus / Q000737:chemistry* / Q000187:drug effects / Q000502:physiology*; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid; D013816:Thermodynamics; D014760:Viral Fusion Proteins / Q000737:chemistry* / Q000378:metabolism*; D010948:Viral Plaque Assay
chemical_list: D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins