Six-helix bundle assembly and characterization of heptad repeat regions from the F protein of Newcastle disease virus.
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.
Yu
Ming
M
Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
Wang
Enxiu
E
Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
Liu
Youfang
Y
Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
Cao
Dianjun
D
Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, People's Republic of China2.
Jin
Ningyi
N
Dept of Virus Research, University of Military Supplies, Changchun 130062, People's Republic of China3.
Zhang
Catherine W-H
CW
Laboratory of Immunobiology, Dana-Farber Cancer Institute and Dept of Medicine, Harvard Medical School, Boston, MA 02115, USA4.
Bartlam
Mark
M
Laboratory of Structural Biology and MOE Laboratory of Protein Sciences, Tsinghua University, Beijing 100084, People's Republic of China5.
Rao
Zihe
Z
Laboratory of Structural Biology and MOE Laboratory of Protein Sciences, Tsinghua University, Beijing 100084, People's Republic of China5.
Tien
Po
P
Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
Gao
George F
GF
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.
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
D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins