Gao Daquan, Narasimhan Diwahar L, Macdonald Joanne, Brim Remy, Ko Mei-Chuan, Landry Donald W, Woods James H, Sunahara Roger K, Zhan Chang-Guo
Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, Kentucky 40536, USA.
Mol Pharmacol. 2009 Feb;75(2):318-23. doi: 10.1124/mol.108.049486. Epub 2008 Nov 5.
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 approximately 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.
通过给予可卡因酯酶（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景的策略，因为CocE是迄今已鉴定出的代谢天然存在的可卡因的最有效天然酶。CocE临床应用的一个主要障碍是天然CocE的热不稳定性，其在生理温度（37摄氏度）下的半衰期仅为几分钟。在此，我们报告了通过使用一种新颖的计算方法进行合理设计，随后进行体外和体内研究而开发出的CocE热稳定变体。这种计算与实验相结合的努力产生了一种CocE变体，其在体外和体内的血浆半衰期均增加了约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突变体。
