Structural analysis of thermostabilizing mutations of cocaine esterase

D L Narasimhan; M R Nance; D Gao; M-C Ko; Joanne Macdonald; P Tamburi; D Yoon; D W Landry; J H Woods; C-G Zhan; J J C Tesmer; R K Sunahara

doi:10.1093/protein/gzq025

Back

Structural analysis of thermostabilizing mutations of cocaine esterase

Journal article

Peer reviewed

Structural analysis of thermostabilizing mutations of cocaine esterase

D L Narasimhan, M R Nance, D Gao, M-C Ko, Joanne Macdonald, P Tamburi, D Yoon, D W Landry, J H Woods, C-G Zhan, …

Protein Engineering Design and Selection, Vol.23(7), pp.537-547

2010

DOI: https://doi.org/10.1093/protein/gzq025

Files and links (1)

url

https://doi.org/10.1093/protein/gzq025View

Published Version

Abstract

cocaine esterase

computational

drug abuse

thermostable

Cocaine is considered to be the most addictive of all substances of abuse and mediates its effects by inhibiting monoamine transporters, primarily the dopamine transporters. There are currently no small molecules that can be used to combat its toxic and addictive properties, in part because of the difficulty of developing compounds that inhibit cocaine binding without having intrinsic effects on dopamine transport. Most of the effective cocaine inhibitors also display addictive properties. We have recently reported the use of cocaine esterase (CocE) to accelerate the removal of systemic cocaine and to prevent cocaine-induced lethality. However, wild-type CocE is relatively unstable at physiological temperatures (τ1/2 ∼13 min at 37°C), presenting challenges for its development as a viable therapeutic agent. We applied computational approaches to predict mutations to stabilize CocE and showed that several of these have increased stability both in vitro and in vivo, with the most efficacious mutant (T172R/G173Q) extending half-life up to 370 min. Here we present novel X-ray crystallographic data on these mutants that provide a plausible model for the observed enhanced stability. We also more extensively characterize the previously reported variants and report on a new stabilizing mutant, L169K. The improved stability of these engineered CocE enzymes will have a profound influence on the use of this protein to combat cocaine-induced toxicity and addiction in humans.

Details

Title: Structural analysis of thermostabilizing mutations of cocaine esterase
Authors: D L Narasimhan (Author) - University of Michigan, United States
M R Nance (Author) - University of Michigan, United States
D Gao (Author) - University of Kentucky, United States
M-C Ko (Author) - University of Michigan, United States
Joanne Macdonald (Author) - Columbia University, United States
P Tamburi (Author) - Columbia University, United States
D Yoon (Author) - University of Michigan, United States
D W Landry (Author) - Columbia University, United States
J H Woods (Author) - University of Michigan, United States
C-G Zhan (Author) - University of Kentucky, United States
J J C Tesmer (Author) - University of Michigan, United States
R K Sunahara (Author) - University of Michigan, United States
Publication details: Protein Engineering Design and Selection, Vol.23(7), pp.537-547
Publisher: Oxford University Press
Date published: 2010
DOI: 10.1093/protein/gzq025
ISSN: 1741-0126
Organisation Unit: School of Science and Engineering - Legacy; University of the Sunshine Coast, Queensland; School of Science, Technology and Engineering; Centre for Bioinnovation
Language: English
Record Identifier: 99450397702621
Output Type: Journal article

Metrics

1 File views/ downloads

882 Record Views

42 Times Cited - Web of Science

InCites Highlights

These are selected metrics from InCites Benchmarking & Analytics tool, related to this output

Collaboration types: Domestic collaboration
Web Of Science research areas: Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology

UN Sustainable Development Goals (SDGs)

This output has contributed to the advancement of the following goals:

Source: InCites