Identification of protein components of vaccinia virus particles
EMSL Project ID
10790
Abstract
Vaccinia virus, the prototype member of the poxvirus family, is predicted to encode nearly 200 proteins. The infectious particle has a complex structure consisting of a lipoprotein membrane surrounding a core. No comprehensive analysis of the protein components of the virus particle has been made. It has been suggested that the virus particle is composed of between 50 and 100 or more polypeptides based on the number of spots on a 2-dimensional gel. We propose to utilize an ultrasensitive nanoscale proteomics approach to identify the protein components of the whole particle. Vaccinia virus will be purified from infected HeLa cells using sucrose gradient centrifugation and solubilized with SDS. The soluble proteins will be digested with trypsin and the peptides prepared for analysis by capillary liquid chromatography coupled to a Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. In subsequent experiments, the virus particle will be separated into surface membrane and core fractions, and the analysis repeated. It is anticipated that this information will be extremely useful for basic research on poxviruses and will aid efforts to make improved vaccines. This work will forward the mission of the NIH in prevention of infectious disease.
Project Details
Project type
Exploratory Research
Start Date
2004-08-27
End Date
2007-08-28
Status
Closed
Released Data Link
Team
Principal Investigator
Related Publications
Protein composition of the vaccinia virus mature virion. Virology. 2007 Feb 5;358(1):233-47.