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Cloning and characterization of a bovine alpha interferon receptor
1Department of Molecular Genetics and Microbiology, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway 08854.
Insights
Researchers isolated a bovine interferon alpha receptor (BoIFN-alpha R1) cDNA. This receptor, when expressed in cells, binds human interferon alpha (IFN-alpha) with high affinity, confirming its functional similarity to the human counterpart.
Area of Science:
- Molecular biology
- Immunology
- Receptor research
Background:
- Interferon alpha (IFN-alpha) plays a crucial role in antiviral responses.
- Understanding interferon receptor function is key to developing antiviral therapies.
- The bovine interferon alpha receptor (BoIFN-alpha R1) had not been fully characterized.
Purpose of the Study:
- To isolate and characterize the bovine interferon alpha receptor (BoIFN-alpha R1) cDNA.
- To determine if the expressed bovine receptor can bind human IFN-alpha.
- To investigate the molecular interactions between human IFN-alpha and the bovine receptor.
Main Methods:
- Isolation of BoIFN-alpha R1 cDNA.
- Transfection of monkey COS cells with the cDNA.
- Binding assays using radiolabeled human IFN-alpha A and IFN-alpha B.
- Covalent crosslinking and SDS-PAGE analysis.
Main Results:
- Successfully isolated a BoIFN-alpha R1 cDNA homologous to the human version.
- Transfected COS cells showed a significant increase in high-affinity binding sites for human IFN-alpha A and B.
- Crosslinking experiments revealed a 140-150 kDa complex formed between radiolabeled human IFN-alpha and the expressed bovine receptor protein.
Conclusions:
- The isolated BoIFN-alpha R1 cDNA encodes a functional receptor capable of binding human IFN-alpha.
- The bovine and human interferon alpha receptors share functional similarities.
- This study provides a basis for further research into interferon receptor interactions and potential therapeutic applications.
Abstract:
A bovine interferon alpha receptor (BoIFN-alpha R1) cDNA, homologous to the human cDNA, was isolated. Transfection of the BoIFN-alpha R1 cDNA into monkey COS cells results in a large increase in high-affinity binding sites for human IFN-alpha A and IFN-alpha B. Covalent crosslinking of radiolabeled HuIFN-alpha A and -alpha B demonstrates that the complex of [32P]HuIFN with the BoIFN-alpha R1 protein (predicted mass, 61,375) expressed in COS cells migrates as a 140-150 kDa band.