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Crystal structure of a bacterial cocaine esterase

Nature Structural Biology volume 9, pages 17–21 (2002)Cite this article

Abstract

Here we report the first structure of a cocaine-degrading enzyme. The bacterial esterase, cocE, hydrolyzes pharmacologically active (−)-cocaine to a nonpsychoactive metabolite with a rate faster than any other reported cocaine esterase (kcat = 7.8 s−1 and KM = 640 nM). Because of the high catalytic proficiency of cocE, it is an attractive candidate for novel protein-based therapies for cocaine overdose. The crystal structure of cocE, solved by multiple anomalous dispersion (MAD) methods, reveals that cocE is a serine esterase composed of three domains: (i) a canonical α/β hydrolase fold (ii) an α-helical domain that caps the active site and (iii) a jelly-roll-like β-domain that interacts extensively with the other two domains. The active site was identified within the interface of all three domains by analysis of the crystal structures of transition state analog adduct and product complexes, which were refined at 1.58 Å and 1.63 Å resolution, respectively. These structural studies suggest that substrate recognition arises partly from interactions between the benzoyl moiety of cocaine and a highly evolved specificity pocket.

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Figure 1: Cocaine hydrolysis.
Figure 2: CocE structure and topology.
Figure 3: Stereo views of the active site of cocE.
Figure 4: Proposed mechanism for acyl intermediate hydrolysis.

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Acknowledgements

The authors thank the Advanced Light Source at Berkeley and the Stanford Synchrotron Radiation Laboratory (SSRL), in particular A. Gonzalez, for advice on MAD data collection. We are also indebted to R. Stanfield for assistance on synchrotron trips and M. Elsliger for helpful discussions. Support was provided by the National Institutes of Health (I.A.W.), the Biotechnology and Biological Sciences Research Council (N.C.B.), the Howard Hughes Medical Institute (J.M.T.), and the Bernie Gilula Foundation (N.A.L.).

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Authors and Affiliations

  1. Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Rd., La Jolla, 92037, California, USA

    Nicholas A. Larsen, James M. Turner, James Stevens, Richard A. Lerner & Ian A. Wilson

  2. Institute of Biotechnology, Cambridge University, Tennis Court Rd., Cambridge, CB2 1QT, UK

    Susan J. Rosser, Amrik Basran & Neil C. Bruce

  3. The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Rd., La Jolla, 92037, California, USA

    Richard A. Lerner & Ian A. Wilson

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  1. Nicholas A. Larsen
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Correspondence to Ian A. Wilson.

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Larsen, N., Turner, J., Stevens, J. et al. Crystal structure of a bacterial cocaine esterase. Nat Struct Mol Biol 9, 17–21 (2002). https://doi.org/10.1038/nsb742

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