Catalytic activities of a cocaine hydrolase engineered from human butyrylcholinesterase against (+)- and (-)-cocaine

Liu Xue; Shurong Hou; Wenchao Yang; Lei Fang; Fang Zheng; Chang Guo Zhan

doi:10.1016/j.cbi.2012.08.003

Catalytic activities of a cocaine hydrolase engineered from human butyrylcholinesterase against (+)- and (-)-cocaine

Liu Xue, Shurong Hou, Wenchao Yang, Lei Fang, Fang Zheng, Chang Guo Zhan

Pharmaceutical Sciences

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

It can be argued that an ideal anti-cocaine medication would be one that accelerates cocaine metabolism producing biologically inactive metabolites via a route similar to the primary cocaine-metabolizing pathway, i.e., hydrolysis catalyzed by butyrylcholinesterase (BChE) in plasma. However, wild-type BChE has a low catalytic efficiency against naturally occurring (-)-cocaine. Interestingly, wild-type BChE has a much higher catalytic activity against unnatural (+)-cocaine. According to available positron emission tomography (PET) imaging analysis using [¹¹C](-)-cocaine and [¹¹C](+)- cocaine tracers in human subjects, only [¹¹C](-)-cocaine was observed in the brain, whereas no significant [¹¹C](+)-cocaine signal was observed in the brain. The available PET data imply that an effective therapeutic enzyme for treatment of cocaine abuse could be an exogenous cocaine-metabolizing enzyme with a catalytic activity against (-)-cocaine comparable to that of wild-type BChE against (+)-cocaine. Our recently designed A199S/F227A/S287G/A328 W/Y332G mutant of human BChE has a considerably improved catalytic efficiency against (-)-cocaine and has been proven active in vivo. In the present study, we have characterized the catalytic activities of wild-type BChE and the A199S/F227A/S287G/A328 W/Y332G mutant against both (+)- and (-)-cocaine at the same time under the same experimental conditions. Based on the obtained kinetic data, the A199S/F227A/S287G/A328 W/Y332G mutant has a similarly high catalytic efficiency (k_cat/K_M) against (+)- and (-)-cocaine, and indeed has a catalytic efficiency (k_cat/K _M = 1.84 × 10⁹ M^-1 min^-1) against (-)-cocaine comparable to that (k_cat/K_M = 1.37 × 10⁹ M^-1 min^-1) of wild-type BChE against (+)-cocaine. Thus, the mutant may be used to effectively prevent (-)-cocaine from entering brain and producing physiological effects in the enzyme-based treatment of cocaine abuse.

Original language	English
Pages (from-to)	57-62
Number of pages	6
Journal	Chemico-Biological Interactions
Volume	203
Issue number	1
DOIs	https://doi.org/10.1016/j.cbi.2012.08.003
State	Published - Mar 25 2013

Bibliographical note

Funding Information:
This work was supported in part by the NIH (Grants R01 DA032910, R01 DA013930, and R01 DA025100 to Zhan) and the NSF (Grant CHE-1111761 to Zhan). The authors also acknowledge the Computer Center at University of Kentucky for supercomputing time on a Dell X-series Cluster with 384 nodes or 4,768 processors.

Funding

This work was supported in part by the NIH (Grants R01 DA032910, R01 DA013930, and R01 DA025100 to Zhan) and the NSF (Grant CHE-1111761 to Zhan). The authors also acknowledge the Computer Center at University of Kentucky for supercomputing time on a Dell X-series Cluster with 384 nodes or 4,768 processors.

Funders	Funder number
National Institutes of Health (NIH)
Author National Institute on Drug Abuse DA031791 Mark J Ferris National Institute on Drug Abuse DA006634 Mark J Ferris National Institute on Alcohol Abuse and Alcoholism AA026117 Mark J Ferris National Institute on Alcohol Abuse and Alcoholism AA028162 Elizabeth G Pitts National Institute of General Medical Sciences GM102773 Elizabeth G Pitts Peter McManus Charitable Trust Mark J Ferris National Institute on Drug Abuse	R01DA013930, R01DA025100, R01DA032910
U.S. Department of Energy Chinese Academy of Sciences Guangzhou Municipal Science and Technology Project Oak Ridge National Laboratory Extreme Science and Engineering Discovery Environment National Science Foundation National Energy Research Scientific Computing Center National Natural Science Foundation of China	CHE-1111761

Keywords

Cholinesterase
Cocaine addiction
Drug overdose
Enzyme therapy
Hydrolase

ASJC Scopus subject areas

Toxicology

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10.1016/j.cbi.2012.08.003

Cite this

@article{143e65d4997d4fb8a2ecb51b5eacd149,

title = "Catalytic activities of a cocaine hydrolase engineered from human butyrylcholinesterase against (+)- and (-)-cocaine",

abstract = "It can be argued that an ideal anti-cocaine medication would be one that accelerates cocaine metabolism producing biologically inactive metabolites via a route similar to the primary cocaine-metabolizing pathway, i.e., hydrolysis catalyzed by butyrylcholinesterase (BChE) in plasma. However, wild-type BChE has a low catalytic efficiency against naturally occurring (-)-cocaine. Interestingly, wild-type BChE has a much higher catalytic activity against unnatural (+)-cocaine. According to available positron emission tomography (PET) imaging analysis using [11C](-)-cocaine and [11C](+)- cocaine tracers in human subjects, only [11C](-)-cocaine was observed in the brain, whereas no significant [11C](+)-cocaine signal was observed in the brain. The available PET data imply that an effective therapeutic enzyme for treatment of cocaine abuse could be an exogenous cocaine-metabolizing enzyme with a catalytic activity against (-)-cocaine comparable to that of wild-type BChE against (+)-cocaine. Our recently designed A199S/F227A/S287G/A328 W/Y332G mutant of human BChE has a considerably improved catalytic efficiency against (-)-cocaine and has been proven active in vivo. In the present study, we have characterized the catalytic activities of wild-type BChE and the A199S/F227A/S287G/A328 W/Y332G mutant against both (+)- and (-)-cocaine at the same time under the same experimental conditions. Based on the obtained kinetic data, the A199S/F227A/S287G/A328 W/Y332G mutant has a similarly high catalytic efficiency (kcat/KM) against (+)- and (-)-cocaine, and indeed has a catalytic efficiency (kcat/K M = 1.84 × 109 M-1 min-1) against (-)-cocaine comparable to that (kcat/KM = 1.37 × 109 M-1 min-1) of wild-type BChE against (+)-cocaine. Thus, the mutant may be used to effectively prevent (-)-cocaine from entering brain and producing physiological effects in the enzyme-based treatment of cocaine abuse.",

keywords = "Cholinesterase, Cocaine addiction, Drug overdose, Enzyme therapy, Hydrolase",

author = "Liu Xue and Shurong Hou and Wenchao Yang and Lei Fang and Fang Zheng and Zhan, \{Chang Guo\}",

note = "Funding Information: This work was supported in part by the NIH (Grants R01 DA032910, R01 DA013930, and R01 DA025100 to Zhan) and the NSF (Grant CHE-1111761 to Zhan). The authors also acknowledge the Computer Center at University of Kentucky for supercomputing time on a Dell X-series Cluster with 384 nodes or 4,768 processors. ",

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T1 - Catalytic activities of a cocaine hydrolase engineered from human butyrylcholinesterase against (+)- and (-)-cocaine

AU - Xue, Liu

AU - Hou, Shurong

AU - Yang, Wenchao

AU - Fang, Lei

AU - Zheng, Fang

AU - Zhan, Chang Guo

N1 - Funding Information: This work was supported in part by the NIH (Grants R01 DA032910, R01 DA013930, and R01 DA025100 to Zhan) and the NSF (Grant CHE-1111761 to Zhan). The authors also acknowledge the Computer Center at University of Kentucky for supercomputing time on a Dell X-series Cluster with 384 nodes or 4,768 processors.

PY - 2013/3/25

Y1 - 2013/3/25

N2 - It can be argued that an ideal anti-cocaine medication would be one that accelerates cocaine metabolism producing biologically inactive metabolites via a route similar to the primary cocaine-metabolizing pathway, i.e., hydrolysis catalyzed by butyrylcholinesterase (BChE) in plasma. However, wild-type BChE has a low catalytic efficiency against naturally occurring (-)-cocaine. Interestingly, wild-type BChE has a much higher catalytic activity against unnatural (+)-cocaine. According to available positron emission tomography (PET) imaging analysis using [11C](-)-cocaine and [11C](+)- cocaine tracers in human subjects, only [11C](-)-cocaine was observed in the brain, whereas no significant [11C](+)-cocaine signal was observed in the brain. The available PET data imply that an effective therapeutic enzyme for treatment of cocaine abuse could be an exogenous cocaine-metabolizing enzyme with a catalytic activity against (-)-cocaine comparable to that of wild-type BChE against (+)-cocaine. Our recently designed A199S/F227A/S287G/A328 W/Y332G mutant of human BChE has a considerably improved catalytic efficiency against (-)-cocaine and has been proven active in vivo. In the present study, we have characterized the catalytic activities of wild-type BChE and the A199S/F227A/S287G/A328 W/Y332G mutant against both (+)- and (-)-cocaine at the same time under the same experimental conditions. Based on the obtained kinetic data, the A199S/F227A/S287G/A328 W/Y332G mutant has a similarly high catalytic efficiency (kcat/KM) against (+)- and (-)-cocaine, and indeed has a catalytic efficiency (kcat/K M = 1.84 × 109 M-1 min-1) against (-)-cocaine comparable to that (kcat/KM = 1.37 × 109 M-1 min-1) of wild-type BChE against (+)-cocaine. Thus, the mutant may be used to effectively prevent (-)-cocaine from entering brain and producing physiological effects in the enzyme-based treatment of cocaine abuse.

AB - It can be argued that an ideal anti-cocaine medication would be one that accelerates cocaine metabolism producing biologically inactive metabolites via a route similar to the primary cocaine-metabolizing pathway, i.e., hydrolysis catalyzed by butyrylcholinesterase (BChE) in plasma. However, wild-type BChE has a low catalytic efficiency against naturally occurring (-)-cocaine. Interestingly, wild-type BChE has a much higher catalytic activity against unnatural (+)-cocaine. According to available positron emission tomography (PET) imaging analysis using [11C](-)-cocaine and [11C](+)- cocaine tracers in human subjects, only [11C](-)-cocaine was observed in the brain, whereas no significant [11C](+)-cocaine signal was observed in the brain. The available PET data imply that an effective therapeutic enzyme for treatment of cocaine abuse could be an exogenous cocaine-metabolizing enzyme with a catalytic activity against (-)-cocaine comparable to that of wild-type BChE against (+)-cocaine. Our recently designed A199S/F227A/S287G/A328 W/Y332G mutant of human BChE has a considerably improved catalytic efficiency against (-)-cocaine and has been proven active in vivo. In the present study, we have characterized the catalytic activities of wild-type BChE and the A199S/F227A/S287G/A328 W/Y332G mutant against both (+)- and (-)-cocaine at the same time under the same experimental conditions. Based on the obtained kinetic data, the A199S/F227A/S287G/A328 W/Y332G mutant has a similarly high catalytic efficiency (kcat/KM) against (+)- and (-)-cocaine, and indeed has a catalytic efficiency (kcat/K M = 1.84 × 109 M-1 min-1) against (-)-cocaine comparable to that (kcat/KM = 1.37 × 109 M-1 min-1) of wild-type BChE against (+)-cocaine. Thus, the mutant may be used to effectively prevent (-)-cocaine from entering brain and producing physiological effects in the enzyme-based treatment of cocaine abuse.

KW - Cholinesterase

KW - Cocaine addiction

KW - Drug overdose

KW - Enzyme therapy

KW - Hydrolase

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Catalytic activities of a cocaine hydrolase engineered from human butyrylcholinesterase against (+)- and (-)-cocaine

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

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