Related Experiment Videos
Processing of pseudorabies virus glycoprotein gII
U Wölfer1, V Kruft, D Sawitzky
1Institut für Klinische und Experimentelle Virologie der Freien Universität Berlin, Federal Republic of Germany.
Journal of Virology
|June 1, 1990
Summary
The pseudorabies virus glycoprotein complex gII, composed of subunits gIIa, gIIb, and gIIc, was analyzed. Results indicate gIIa is a minor component, while gIIb and gIIc are equimolar fragments derived from gIIa.
Area of Science:
- Virology
- Molecular Biology
- Protein Chemistry
Background:
- The pseudorabies virus (PRV) glycoprotein complex gII is crucial for viral infection.
- Understanding the subunit composition and processing of gII is essential for viral pathogenesis research.
Purpose of the Study:
- To elucidate the subunit composition and processing of the PRV glycoprotein complex gII.
- To determine the N-terminal sequences and relative stoichiometry of gII subunits.
Main Methods:
- Immunoprecipitation of PRV gII using monoclonal antibody M5 and protein A-Sepharose.
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions.
- N-terminal sequencing of isolated subunits and protein labeling with 14C-amino acids.
Main Results:
- The glycoprotein complex gII consists of subunits gIIa, gIIb, and gIIc.
- N-terminal sequencing revealed gIIa and gIIb share a common start site (position 59), while gIIc starts later (position 503).
- Protein labeling indicated gIIa is a minor component, whereas gIIb and gIIc are present in equimolar amounts, suggesting gIIa is cleaved into gIIb and gIIc.
Conclusions:
- The PRV glycoprotein gII undergoes post-translational cleavage.
- Subunits gIIb and gIIc represent equimolar fragments derived from a larger precursor, likely initiated as gIIa.
- The stoichiometry suggests a specific processing pathway for the PRV gII complex.