Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

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Kirkpatrick, R.B. et al.

Kirkpatrick, Grooms, Wang, Fenderson, Feild, Pratta, Volker, Scott, Johanson,

Bacterial production of biologically active canine interleukin-1beta by seamless SUMO tagging and removal.

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 chondroctyes. This method combines highly efficient aspects of seamless plasmid engineering, protein purification, and precise tag removal.[1]

References

Bacterial production of biologically active canine interleukin-1beta by seamless SUMO tagging and removal. Kirkpatrick, R.B., Grooms, M., Wang, F., Fenderson, H., Feild, J., Pratta, M.A., Volker, C., Scott, G., Johanson, K. Protein Expr. Purif. (2006) [Pubmed]

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