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A screening system for the identification of refolding conditions for a model protein kinase, p38 alpha
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Cowan, Richard H., Davies, Rick A. and Pinheiro, Teresa J. T. (2008) A screening system for the identification of refolding conditions for a model protein kinase, p38 alpha. Analytical Biochemistry, Vol.376 (No.1). pp. 25-38. doi:10.1016/j.ab.2008.01.036 ISSN 0003-2697.
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Official URL: http://dx.doi.org/10.1016/j.ab.2008.01.036
Abstract
Protein kinases are key drug targets involved in the regulation of a wide variety of cellular processes. To aid the development of drugs targeting these kinases, it is necessary to express recombinant protein in large amounts. The expression of these kinases in Escherichia coli often leads to the accumulation of the expressed protein as insoluble inclusion bodies. The refolding of these inclusion bodies could provide a route to soluble protein, but there is little reported success in this area. We set out to develop a system for the screening of refolding conditions for a model protein kinase, p38 alpha, and applied this system to denatured p38 alpha derived from natively folded and inclusion body protein. Clear differences were observed in the refolding yields obtained, suggesting differences in the folded state of these preparations. Using the screening system, we have established conditions under which soluble, folded p38 alpha can be produced from inclusion bodies. We have shown that the refolding yields obtained in this screen are suitable for the economic large-scale production of refolded p38 alpha protein kinase. (c) 2008 Elsevier Inc. All rights reserved.
| Item Type: | Journal Article | ||||
|---|---|---|---|---|---|
| Subjects: | Q Science > QP Physiology | ||||
| Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) > Biological Sciences ( -2010) | ||||
| Library of Congress Subject Headings (LCSH): | Protein kinases, Protein folding, Recombinant proteins | ||||
| Journal or Publication Title: | Analytical Biochemistry | ||||
| Publisher: | Academic Press | ||||
| ISSN: | 0003-2697 | ||||
| Official Date: | 1 May 2008 | ||||
| Dates: |
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| Volume: | Vol.376 | ||||
| Number: | No.1 | ||||
| Number of Pages: | 14 | ||||
| Page Range: | pp. 25-38 | ||||
| DOI: | 10.1016/j.ab.2008.01.036 | ||||
| Status: | Peer Reviewed | ||||
| Publication Status: | Published | ||||
| Access rights to Published version: | Restricted or Subscription Access | ||||
| Funder: | Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), AstraZeneca (Firm) |
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