Abstract
Enhancing cocaine metabolism by administration of cocaine esterase (CocE) has been recognized as a promising treatment strategy for cocaine overdose and addiction, because CocE is the most efficient native enzyme for metabolizing the naturally occurring cocaine yet identified. A major obstacle to the clinical application of CocE is the thermoinstability of native CocE with a half-life of only a few minutes at physiological temperature (37°C). Here we report thermostable variants of CocE developed through rational design using a novel computational approach followed by in vitro and in vivo studies. This integrated computational-experimental effort has yielded a CocE variant with a ∼30-fold increase in plasma half-life both in vitro and in vivo. The novel design strategy can be used to develop thermostable mutants of any protein.
Footnotes
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This work was supported by the National Institutes of Health [Grants DA021416, DA025100, DA013930, and GM007767].
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D.G. and D.L.N. contributed equally to this work.
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ABBREVIATIONS: CocE, cocaine esterase; MD, molecular dynamics; RMSD, root-mean-square deviation; wt, wild type; PAGE, polyacrylamide gel electrophoresis.
- Received June 5, 2008.
- Accepted November 4, 2008.
- The American Society for Pharmacology and Experimental Therapeutics
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