Thermostable Variants of Cocaine Esterase for Long-Time Protection against Cocaine Toxicity
Thermostable Variants of Cocaine Esterase for Long-Time Protection against Cocaine Toxicity
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DOI：
10.1124/mol.108.049486
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发表时间：
2009-02-01
影响因子：
3.6
通讯作者：
Zhan, Chang-Guo
中科院分区：
文献类型：
--
作者：
Gao, Daquan;Narasimhan, Diwahar L.;Zhan, Chang-Guo
通过施用可卡因酯酶 (CocE) 来增强可卡因代谢已被认为是治疗可卡因过量和成瘾的一种有前途的治疗策略，因为 CocE 是迄今为止已确定的代谢天然可卡因的最有效的天然酶。 CocE临床应用的一个主要障碍是天然CocE的热稳定性，在生理温度（37℃）下半衰期仅为几分钟。在这里，我们报告了使用新颖的计算方法进行合理设计并随后进行体外和体内研究的 CocE 热稳定变体。这种综合计算实验的努力已经产生了一种 CocE 变体，其血浆半衰期在体外和体内均增加了 30 倍。这种新颖的设计策略可用于开发任何蛋白质的热稳定性突变体。
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 degrees 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 similar to 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.