TY - JOUR
T1 - Thermostable variants of cocaine esterase for long-time protection against cocaine toxicity
AU - Gao, Daquan
AU - Narasimhan, Diwahar L.
AU - Macdonald, Joanne
AU - Brim, Remy
AU - Ko, Mei Chuan
AU - Landry, Donald W.
AU - Woods, James H.
AU - Sunahara, Roger K.
AU - Zhan, Chang Guo
PY - 2009/2
Y1 - 2009/2
N2 - 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.
AB - 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.
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U2 - 10.1124/mol.108.049486
DO - 10.1124/mol.108.049486
M3 - Article
C2 - 18987161
AN - SCOPUS:59449089478
SN - 0026-895X
VL - 75
SP - 318
EP - 323
JO - Molecular Pharmacology
JF - Molecular Pharmacology
IS - 2
ER -