Thermostable variants of cocaine esterase for long-time protection against cocaine toxicity.
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.
Gao
Daquan
D
Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, Kentucky 40536, USA.
Narasimhan
Diwahar L
DL
Macdonald
Joanne
J
Brim
Remy
R
Ko
Mei-Chuan
MC
Landry
Donald W
DW
Woods
James H
JH
Sunahara
Roger K
RK
Zhan
Chang-Guo
CG
D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism / Q000627:therapeutic use*; D003042:Cocaine / Q000008:administration & dosage / Q000378:metabolism* / Q000633:toxicity; D019970:Cocaine-Related Disorders / Q000150:complications / Q000188:drug therapy*; D004195:Disease Models, Animal; D015195:Drug Design; D062787:Drug Overdose; D008297:Male; D051379:Mice; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D002265:Carboxylic Ester Hydrolases; C019921:cocaine esterase; D003042:Cocaine