3IDA

Thermostable Cocaine Esterase with mutations L169K and G173Q, bound to DTT adduct

Summary for 3IDA

• Entry DOI: 10.2210/pdb3ida/pdb
• Related: 3I2I 3i2g 3i2j 3i2k
• Descriptor: Cocaine esterase, CHLORIDE ION, (4S,5S)-4,5-BIS(MERCAPTOMETHYL)-1,3-DIOXOLAN-2-OL, ... (5 entities in total)
• Functional Keywords: alpha/beta hydrolase, esterase, hydrolase
• Biological source: Rhodococcus sp.
• Cellular location: Cytoplasm (Probable): Q9L9D7
• Total number of polymer chains: 1
• Total formula weight: 64822.57

Authors

Tesmer, J.J.G.,Nance, M.R. (deposition date: 2009-07-20, release date: 2010-03-02, Last modification date: 2023-09-06)

Primary citation

Brim, R.L.,Nance, M.R.,Youngstrom, D.W.,Narasimhan, D.,Zhan, C.G.,Tesmer, J.J.,Sunahara, R.K.,Woods, J.H.
A thermally stable form of bacterial cocaine esterase: a potential therapeutic agent for treatment of cocaine abuse.
Mol.Pharmacol., 77:593-600, 2010

PubMed Abstract: Rhodococcal cocaine esterase (CocE) is an attractive potential treatment for both cocaine overdose and cocaine addiction. CocE directly degrades cocaine into inactive products, whereas traditional small-molecule approaches require blockade of the inhibitory action of cocaine on a diverse array of monoamine transporters and ion channels. The usefulness of wild-type (wt) cocaine esterase is hampered by its inactivation at 37 degrees C. Herein, we characterize the most thermostable form of this enzyme to date, CocE-L169K/G173Q. In vitro kinetic analyses reveal that CocE-L169K/G173Q displays a half-life of 2.9 days at 37 degrees C, which represents a 340-fold improvement over wt and is 15-fold greater than previously reported mutants. Crystallographic analyses of CocE-L169K/G173Q, determined at 1.6-A resolution, suggest that stabilization involves enhanced domain-domain interactions involving van der Waals interactions and hydrogen bonding. In vivo rodent studies reveal that intravenous pretreatment with CocE-L169K/G173Q in mice provides protection from cocaine-induced lethality for longer time periods before cocaine administration than wt CocE. Furthermore, intravenous administration (pretreatment) of CocE-L169K/G173Q prevents self-administration of cocaine in a time-dependent manner. Termination of the in vivo effects of CoCE seems to be dependent on, but not proportional to, its clearance from plasma as its half-life is approximately 2.3 h and similar to that of wt CocE (2.2 h). Taken together these data suggest that CocE-L169K/G173Q possesses many of the properties of a biological therapeutic for treating cocaine abuse but requires additional development to improve its serum half-life.

PubMed: 20086035
DOI: 10.1124/mol.109.060806

Experimental method

X-RAY DIFFRACTION (1.6 Å)