Plant-expressed Cocaine Hydrolase Variants of Butyrylcholinesterase Exhibit Altered Allosteric Effects of Cholinesterase Activity and Increased Inhibitor Sensitivity

Overview

Journal Sci Rep

Specialty Science

Date 2017 Sep 7

PMID 28874829

Citations 13

Authors

Katherine E Larrimore

I Can Kazan

Latha Kannan

R Player Kendle

Tameem Jamal

Matthew Barcus

Ashini Bolia

Stephen Brimijoin

Chang-Guo Zhan

S Banu Ozkan

Tsafrir S Mor

Affiliations

Soon will be listed here.

Abstract

Butyrylcholinesterase (BChE) is an enzyme with broad substrate and ligand specificities and may function as a generalized bioscavenger by binding and/or hydrolyzing various xenobiotic agents and toxicants, many of which target the central and peripheral nervous systems. Variants of BChE were rationally designed to increase the enzyme's ability to hydrolyze the psychoactive enantiomer of cocaine. These variants were cloned, and then expressed using the magnICON transient expression system in plants and their enzymatic properties were investigated. In particular, we explored the effects that these site-directed mutations have over the enzyme kinetics with various substrates of BChE. We further compared the affinity of various anticholinesterases including organophosphorous nerve agents and pesticides toward these BChE variants relative to the wild type enzyme. In addition to serving as a therapy for cocaine addiction-related diseases, enhanced bioscavenging against other harmful agents could add to the practicality and versatility of the plant-derived recombinant enzyme as a multivalent therapeutic.

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References

Kim H, Zou T, Modi C, Dorner K, Grunkemeyer T, Chen L . A hinge migration mechanism unlocks the evolution of green-to-red photoconversion in GFP-like proteins. Structure. 2015; 23(1):34-43. PMC: 4370283. DOI: 10.1016/j.str.2014.11.011. View

Kumar A, Glembo T, Ozkan S . The Role of Conformational Dynamics and Allostery in the Disease Development of Human Ferritin. Biophys J. 2015; 109(6):1273-81. PMC: 4576160. DOI: 10.1016/j.bpj.2015.06.060. View

Saxena A, Hastings N, Sun W, Dabisch P, Hulet S, Jakubowski E . Prophylaxis with human serum butyrylcholinesterase protects Göttingen minipigs exposed to a lethal high-dose of sarin vapor. Chem Biol Interact. 2015; 238:161-9. DOI: 10.1016/j.cbi.2015.07.001. View

Geyer B, Muralidharan M, Cherni I, Doran J, Fletcher S, Evron T . Purification of transgenic plant-derived recombinant human acetylcholinesterase-R. Chem Biol Interact. 2005; 157-158:331-4. DOI: 10.1016/j.cbi.2005.10.097. View

Carmona G, Jufer R, Goldberg S, Gorelick D, Greig N, Yu Q . Butyrylcholinesterase accelerates cocaine metabolism: in vitro and in vivo effects in nonhuman primates and humans. Drug Metab Dispos. 2000; 28(3):367-71. View

Gerek Z, Ozkan S . Change in allosteric network affects binding affinities of PDZ domains: analysis through perturbation response scanning. PLoS Comput Biol. 2011; 7(10):e1002154. PMC: 3188487. DOI: 10.1371/journal.pcbi.1002154. View

Chen X, Fang L, Liu J, Zhan C . Reaction pathway and free energy profiles for butyrylcholinesterase-catalyzed hydrolysis of acetylthiocholine. Biochemistry. 2012; 51(6):1297-305. PMC: 3292049. DOI: 10.1021/bi201786s. View

Schneider J, Marillonnet S, Castilho A, Gruber C, Werner S, Mach L . Oligomerization status influences subcellular deposition and glycosylation of recombinant butyrylcholinesterase in Nicotiana benthamiana. Plant Biotechnol J. 2014; 12(7):832-9. PMC: 4265266. DOI: 10.1111/pbi.12184. View

Atilgan C, Atilgan A . Perturbation-response scanning reveals ligand entry-exit mechanisms of ferric binding protein. PLoS Comput Biol. 2009; 5(10):e1000544. PMC: 2758672. DOI: 10.1371/journal.pcbi.1000544. View

10.

Felder C, Harel M, Silman I, Sussman J . Structure of a complex of the potent and specific inhibitor BW284C51 with Torpedo californica acetylcholinesterase. Acta Crystallogr D Biol Crystallogr. 2002; 58(Pt 10 Pt 2):1765-71. DOI: 10.1107/s0907444902011642. View

11.

Geyer B, Fletcher S, Griffin T, Lopker M, Soreq H, Mor T . Translational control of recombinant human acetylcholinesterase accumulation in plants. BMC Biotechnol. 2007; 7:27. PMC: 1913049. DOI: 10.1186/1472-6750-7-27. View

12.

Geyer B, Kannan L, Garnaud P, Broomfield C, Cadieux C, Cherni I . Plant-derived human butyrylcholinesterase, but not an organophosphorous-compound hydrolyzing variant thereof, protects rodents against nerve agents. Proc Natl Acad Sci U S A. 2010; 107(47):20251-6. PMC: 2996644. DOI: 10.1073/pnas.1009021107. View

13.

Masson P, Xie W, Froment M, Lockridge O . Effects of mutations of active site residues and amino acids interacting with the Omega loop on substrate activation of butyrylcholinesterase. Biochim Biophys Acta. 2001; 1544(1-2):166-76. DOI: 10.1016/s0167-4838(00)00217-x. View

14.

Xue L, Hou S, Tong M, Fang L, Chen X, Jin Z . Preparation and in vivo characterization of a cocaine hydrolase engineered from human butyrylcholinesterase for metabolizing cocaine. Biochem J. 2013; 453(3):447-54. PMC: 4365901. DOI: 10.1042/BJ20130549. View

15.

Chen X, Huang X, Geng L, Xue L, Hou S, Zheng X . Kinetic characterization of a cocaine hydrolase engineered from mouse butyrylcholinesterase. Biochem J. 2014; 466(2):243-51. PMC: 4367957. DOI: 10.1042/BJ20141266. View

16.

Xue L, Hou S, Yang W, Fang L, Zheng F, Zhan C . Catalytic activities of a cocaine hydrolase engineered from human butyrylcholinesterase against (+)- and (-)-cocaine. Chem Biol Interact. 2012; 203(1):57-62. PMC: 3527670. DOI: 10.1016/j.cbi.2012.08.003. View

17.

Barak D, Ordentlich A, Bromberg A, Kronman C, Marcus D, Lazar A . Allosteric modulation of acetylcholinesterase activity by peripheral ligands involves a conformational transition of the anionic subsite. Biochemistry. 1995; 34(47):15444-52. DOI: 10.1021/bi00047a008. View

18.

Boeck A, Schopfer L, Lockridge O . DNA sequence of butyrylcholinesterase from the rat: expression of the protein and characterization of the properties of rat butyrylcholinesterase. Biochem Pharmacol. 2002; 63(12):2101-10. DOI: 10.1016/s0006-2952(02)01029-8. View

19.

Atilgan A, Durell S, Jernigan R, Demirel M, Keskin O, Bahar I . Anisotropy of fluctuation dynamics of proteins with an elastic network model. Biophys J. 2001; 80(1):505-15. PMC: 1301252. DOI: 10.1016/S0006-3495(01)76033-X. View

20.

Gao D, Zhan C . Modeling evolution of hydrogen bonding and stabilization of transition states in the process of cocaine hydrolysis catalyzed by human butyrylcholinesterase. Proteins. 2005; 62(1):99-110. PMC: 2882100. DOI: 10.1002/prot.20713. View