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.
MeSH Terms
Amino Acid Sequence
Cell Membrane/drug effects,metabolism,virology
China
Circular Dichroism
HeLa Cells
Humans
Inhibitory Concentration 50
Membrane Fusion/drug effects
Models, Molecular
Molecular Sequence Data
Molecular Weight
Newcastle disease virus/chemistry,drug effects,physiology
Peptide Fragments/chemistry,pharmacology
Protein Structure, Secondary
Protein Structure, Tertiary
Recombinant Fusion Proteins/chemistry,metabolism
Repetitive Sequences, Amino Acid
Thermodynamics
Viral Fusion Proteins/chemistry,metabolism
Viral Plaque Assay
Chemicals
Peptide Fragments
Recombinant Fusion Proteins
Viral Fusion Proteins
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Yu Ming
Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
Wang Enxiu
Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
Liu Youfang
Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
Cao Dianjun
Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, People's Republic of China2.
Jin Ningyi
Dept of Virus Research, University of Military Supplies, Changchun 130062, People's Republic of China3.
Zhang Catherine W-H
Laboratory of Immunobiology, Dana-Farber Cancer Institute and Dept of Medicine, Harvard Medical School, Boston, MA 02115, USA4.
Bartlam Mark
Laboratory of Structural Biology and MOE Laboratory of Protein Sciences, Tsinghua University, Beijing 100084, People's Republic of China5.
Rao Zihe
Laboratory of Structural Biology and MOE Laboratory of Protein Sciences, Tsinghua University, Beijing 100084, People's Republic of China5.
Tien Po
Dept of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China1.
Gao George F
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.
