Structural characterization and membrane binding properties of the matrix protein VP40 of Ebola virus.
The matrix protein VP40 of Ebola virus is believed to play a central role in viral assembly as it targets the plasma membrane of infected cells and subsequently forms a tightly packed layer on the inner side of the viral envelope. Expression of VP40 in Escherichia coli and subsequent proteolysis yielded two structural variants differing by a C-terminal truncation 114 amino acid residues long. As indicated by chemical cross-linking studies and electron microscopy, the larger polypeptide was present in a monomeric form, whereas the truncated one formed hexamers. When analyzed for their in vitro binding properties, both constructs showed that only monomeric VP40 efficiently associated with membranes containing negatively charged lipids. Membrane association of truncated, hexameric VP40 was inefficient, indicating a membrane-recognition role for the C-terminal part. Based on these observations we propose that assembly of Ebola virus involves the formation of VP40 hexamers that is mediated by the N-terminal part of the polypeptide.
Ruigrok
R W
RW
Grenoble Outstation, European Molecular Biology Laboratory (EMBL), 6 rue Jules Horowitz, Grenoble, 38000, France.
Schoehn
G
G
Dessen
A
A
Forest
E
E
Volchkov
V
V
Dolnik
O
O
Klenk
H D
HD
Weissenhorn
W
W
D000595:Amino Acid Sequence; D001665:Binding Sites; D003432:Cross-Linking Reagents / Q000378:metabolism; D029043:Ebolavirus / Q000737:chemistry* / Q000502:physiology / Q000648:ultrastructure; D008051:Lipid Bilayers / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism*; D008854:Microscopy, Electron; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009698:Nucleoproteins / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D055672:Static Electricity; D014758:Viral Core Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D019065:Virus Assembly
D003432:Cross-Linking Reagents; D008051:Lipid Bilayers; D008081:Liposomes; D009698:Nucleoproteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; C084819:nucleoprotein VP40, Ebola virus