Bacterial production of biologically active canine interleukin-1β by seamless SUMO tagging and removal
| Autor: | Feilan Wang, Kyung O. Johanson, Heather Fenderson, Robert B. Kirkpatrick, Gilbert F. Scott, Michael A. Pratta, Craig Volker, Michael Grooms, John A. Feild |
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| Rok vydání: | 2006 |
| Předmět: |
DNA
Complementary Interleukin-1beta Molecular Sequence Data Biology medicine.disease_cause law.invention Dogs Plasmid law Complementary DNA Protein purification Escherichia coli medicine Animals Humans Amino Acid Sequence Cloning Molecular Aggrecanase Biological activity Molecular biology Recombinant Proteins Cell biology Gene Expression Regulation Small Ubiquitin-Related Modifier Proteins Recombinant DNA Cattle Extracellular Matrix Degradation Biotechnology |
| Zdroj: | Protein Expression and Purification. 50:102-110 |
| ISSN: | 1046-5928 |
| DOI: | 10.1016/j.pep.2006.05.022 |
| Popis: | Interleukin 1beta (IL-1beta) is a potent stimulator of extracellular matrix degradation in models of osteoarthritis (OA). In contrast to bovine explant models which effectively respond to recombinant human IL-1beta, canine models are relatively refractory to human IL-1beta stimulation. Canine IL-1beta cDNA was cloned in order to produce a fully potent species matched preparation of IL-1beta for use specifically in canine models of OA. Established methods for the production of various orthologous IL-1beta proteins from different species are problematic due to the exquisite sensitivity of the mature IL-1beta product to N-terminal variations and the intrinsic technical challenges associated with producing an unmodified product. We have applied a seamless method of SUMO tagging and removal in order to produce a homogeneous unmodified preparation of canine IL-1beta from Escherichia coli which was found to be a potent inducer of aggrecanase activity in isolated canine articular chondrocytes. This method combines highly efficient aspects of seamless plasmid engineering, protein purification, and precise tag removal. |
| Databáze: | OpenAIRE |
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