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Production of sumoylated proteins using a baculovirus expression system
Martijn A Langereis1, Germán Rosas-Acosta, Klaas Mulder
1Department of Microbial and Molecular Pathogenesis, College of Medicine, Texas A&M Health Science Center, College Station, TX 77843-1114, USA.
Journal of Virological Methods
|January 9, 2007
Summary
Insect cells (Sf9) can be engineered for efficient protein SUMOylation. Introducing mammalian Ubc9 and SUMO proteins enables post-translational modification of exogenous proteins, aiding in the production of sumoylated mammalian proteins.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Spodoptera frugiperda Sf9 cells possess an endogenous sumoylation system.
- The native sumoylation machinery in Sf9 cells is inefficient for modifying exogenous proteins expressed via recombinant baculoviruses.
Purpose of the Study:
- To enhance the sumoylation efficiency of exogenous proteins in Sf9 cells.
- To establish a system for producing sumoylated mammalian proteins in a eukaryotic host.
Main Methods:
- Co-infection of Sf9 cells with recombinant baculoviruses encoding mammalian sumoylation components (Ubc9 and SUMO1 or SUMO3).
- Assessment of sumoylation activity on co-infected test proteins.
Main Results:
- Co-expression of mammalian Ubc9 and SUMO (SUMO1 or SUMO3) was sufficient to confer active sumoylation of exogenous proteins.
- The developed system efficiently modifies test proteins in a eukaryotic environment.
Conclusions:
- Mammalian sumoylation machinery components can be successfully introduced into Sf9 cells to overcome limitations in modifying exogenous proteins.
- This engineered system offers a convenient method for producing sumoylated mammalian proteins, potentially in large quantities for purification.
