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Proteolytic dissection of Sindbis virus core protein
1Harvard University, Cambridge, Massachusetts.
Journal of Virology
|August 1, 1990
Summary
Mild trypsin treatment revealed a stable 18.5 kDa fragment of the Sindbis virus nucleocapsid protein. This finding supports a model of alphavirus core proteins with a flexible N-terminal arm and a compact C-terminal domain.
Area of Science:
- Virology
- Structural Biology
- Protein Chemistry
Background:
- The Sindbis virus nucleocapsid protein is crucial for viral assembly and RNA packaging.
- Previous sequence analysis suggested a domain organization within the alphavirus core proteins.
Purpose of the Study:
- To biochemically characterize the structural domains of the Sindbis virus nucleocapsid protein.
- To provide experimental evidence supporting a proposed model of alphavirus core protein structure.
Main Methods:
- Mild trypsin digestion of purified Sindbis virus nucleocapsid protein.
- Analysis of digestion products using gel exclusion chromatography to determine molecular mass and quaternary structure.
Main Results:
- A stable fragment of approximately 18.5 kilodaltons was generated, N-terminally truncated at residue 105.
- This fragment was found to be monomeric by gel exclusion chromatography.
- The fragment's stability and size are consistent with a compactly folded C-terminal domain.
Conclusions:
- The experimental data support a model where Sindbis virus nucleocapsid protein comprises a flexible N-terminal arm (residues 1-103) and a stable, monomeric C-terminal domain (residues 104-274).
- This domain organization is likely conserved across alphaviruses.
- The findings contribute to understanding the structural basis of alphavirus replication and assembly.