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Structure of an Insecticide Sequestering Carboxylesterase from the Disease Vector Culex quinquefasciatus: What Makes an Enzyme a Good Insecticide Sponge?

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Research School of Chemistry, Australian National University, Canberra, Australian Capital Territory 0200, Australia
CSIRO, GPO Box 1700, Canberra, Australian Capital Territory 2601, Australia
Cite this: Biochemistry 2017, 56, 41, 5512–5525
Publication Date (Web):September 20, 2017
Copyright © 2017 American Chemical Society

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    Abstract Image

    Carboxylesterase (CBE)-mediated metabolic resistance to organophosphate and carbamate insecticides is a major problem for the control of insect disease vectors, such as the mosquito. The most common mechanism involves overexpression of CBEs that bind to the insecticide with high affinity, thereby sequestering them before they can interact with their target. However, the absence of any structure for an insecticide-sequestering CBE limits our understanding of the molecular basis for this process. We present the first structure of a CBE involved in sequestration, Cqestβ21, from the mosquito disease vector Culex quinquefasciatus. Lysine methylation was used to obtain the crystal structure of Cqestβ21, which adopts a canonical α/β-hydrolase fold that has high similarity to the target of organophosphate and carbamate insecticides, acetylcholinesterase. Sequence similarity networks of the insect carboxyl/cholinesterase family demonstrate that CBEs associated with metabolic insecticide resistance across many species share a level of similarity that distinguishes them from a variety of other classes. This is further emphasized by the structural similarities and differences in the binding pocket and active site residues of Cqestβ21 and other insect carboxyl/cholinesterases. Stopped-flow and steady-state inhibition studies support a major role for Cqestβ21 in organophosphate resistance and a minor role in carbamate resistance. Comparison with another isoform associated with insecticide resistance, Cqestβ1, showed both enzymes have similar affinity to insecticides, despite 16 amino acid differences between the two proteins. This provides a molecular understanding of pesticide sequestration by insect CBEs and could facilitate the design of CBE-specific inhibitors to circumvent this resistance mechanism in the future.

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    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.biochem.7b00774.

    • A reaction scheme for LcαE7 G137D, Cqestβ21, and AChE with OP insecticide showing the different paths, a list of NCBI Fasta files included in the SSN to aid group classification, a list of the UniProt IDs of insecticide resistance CBEs used in the SSNs, a table of the chemical structures mentioned in the results and discussion, a multiple sequence alignment of B2 isoforms from the Culex assemblage including a list of Genbank accession numbers (PDF)

    Accession Codes

    PDB ID: 5W1U; PDB ID: 4FG5; PDB ID: 1QO9; PDB ID: 4FNM; PDB ID: 2FJ0; PDB ID: 5THM; Genbank accession number: CAA83643; Genbank accession number: AAA28289.1.

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    Cited By

    This article is cited by 22 publications.

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    13. Annie J. Krueger, Leslie C. Rault, Emily Robinson, Thomas J. Weissling, Ana M. Velez, Troy Anderson. Pyrethroid Insecticide and Milkweed Cardenolide Interactions On Detoxification Enzyme Activity and Expression in Monarch Caterpillars. SSRN Electronic Journal 2022, 65
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    20. Li-sha Bai, Cai-xia Zhao, Jing-jing Xu, Chuan Feng, Yong-qiang Li, Yan-ling Dong, Zhi-qing Ma. Identification and biochemical characterization of carboxylesterase 001G associated with insecticide detoxification in Helicoverpa armigera. Pesticide Biochemistry and Physiology 2019, 157 , 69-79.
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    22. André Q. Torres, Denise Valle, Rafael D. Mesquita, Renata Schama. Gene Family Evolution and the Problem of a Functional Classification of Insect Carboxylesterases. 2018

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