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A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells
Published on: July 31, 2012
Expression and reconstitution of a biologically active human interferon-gamma receptor in hamster cells
1Department of Molecular Genetics and Microbiology, University of Medicine and Dentistry, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854-5635.
Insights
We achieved stable expression and biological activity of the human interferon-gamma (Hu-IFN-gamma) receptor in animal cells. This demonstrates the need for human chromosome 21 and other factors for receptor function.
Area of Science:
- Molecular Biology
- Immunology
- Cell Biology
Background:
- The human interferon-gamma (Hu-IFN-gamma) receptor mediates cellular responses to IFN-gamma.
- Understanding receptor function is crucial for immunology and molecular biology.
Purpose of the Study:
- To achieve stable expression and demonstrate biological activity of the Hu-IFN-gamma receptor in animal cells.
- To investigate the species-specific factors required for Hu-IFN-gamma receptor modulation.
Main Methods:
- Cloning and stable expression of the Hu-IFN-gamma receptor cDNA in animal cells.
- Assessing biological activity through surface histocompatibility antigen expression.
- Utilizing cell lines with and without human chromosome 21.
Main Results:
- Successful stable expression and demonstrated biological activity of the Hu-IFN-gamma receptor.
- Biological activity requires human chromosome 21, indicating a need for additional species-specific factors.
- Established a novel system for studying IFN-gamma receptor function.
Conclusions:
- The Hu-IFN-gamma receptor's biological activity is dependent on species-specific factors, including those on human chromosome 21.
- This expression system facilitates further research into IFN-gamma receptor binding and signal transduction pathways.
Abstract:
We report the expression of the human interferon-gamma (Hu-IFN-gamma) receptor from a cDNA clone in an animal cell where both stable expression of the cloned receptor as well as its biological activity is demonstrated for the first time. Biological activity of the receptor (i.e. expression of surface histocompatibility antigens in response to human interferon-gamma) requires the presence of human chromosome 21 demonstrating the requirement for at least one other species-specific factor in the modulation of receptor action. This system should now facilitate delineation of regions involved in the binding of human interferon-gamma and receptor signal transduction.
