Structure and function of an insect α-carboxylesterase (αEsterase7) associated with insecticide resistance
| Autor: | Paul D. Carr, Faisal Younus, Jian-Wei Liu, Robyn J. Russell, Chris M. Coppin, Tamara Meirelles, John G. Oakeshott, David L. Ollis, Mathilde Lethier, Martin Weik, Gunjan Pandey, Colin J. Jackson |
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| Přispěvatelé: | Research School of Chemistry, Australian National University (ANU), Unit for Virus Host-Cell Interactions [Grenoble] (UVHCI), Centre National de la Recherche Scientifique (CNRS)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Université Joseph Fourier - Grenoble 1 (UJF), CSIRO - Land & Water National Research Flagship, Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Commonwealth Scientific and Industrial Research Organisation [Canberra] (CSIRO), Université Joseph Fourier - Grenoble 1 (UJF)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA) |
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
0106 biological sciences
Insecticides Drug Resistance MESH: Protein Structure Secondary MESH: Catalytic Domain Genes Insect MESH: Genes Insect Crystallography X-Ray 01 natural sciences Protein Structure Secondary Carboxylesterase Substrate Specificity chemistry.chemical_compound Catalytic Domain MESH: Animals MESH: Carboxylesterase Phosphorylation directed evolution MESH: Diptera chemistry.chemical_classification 0303 health sciences Multidisciplinary biology [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] MESH: Sheep Diseases Organophosphate Biological Sciences Directed evolution Acetylcholinesterase Biochemistry Lucilia cuprina MESH: Drug Resistance ali-esterase MESH: Australia Sheep Diseases MESH: Sheep 03 medical and health sciences Animals 030304 developmental biology Enzyme substrate complex Sheep MESH: Phosphorylation Diptera Australia Active site protein engineering MESH: Acetylcholinesterase biology.organism_classification MESH: Crystallography X-Ray MESH: Insecticides 010602 entomology Enzyme chemistry biology.protein MESH: Substrate Specificity |
| Zdroj: | 'Proceedings of the National Academy of Sciences of the USA ', vol: 110, pages: 10177-10182 (2013) Proceedings of the National Academy of Sciences of the United States of America Proceedings of the National Academy of Sciences of the United States of America, National Academy of Sciences, 2013, 110 (25), pp.10177-10182. ⟨10.1073/pnas.1304097110⟩ Proceedings of the National Academy of Sciences of the United States of America, 2013, 110 (25), pp.10177-10182. ⟨10.1073/pnas.1304097110⟩ |
| ISSN: | 0027-8424 1091-6490 |
| DOI: | 10.1073/pnas.1304097110⟩ |
| Popis: | Insect carboxylesterases from the α Esterase gene cluster, such as αE7 (also known as E3) from the Australian sheep blowfly Lucilia cuprina ( Lc αE7), play an important physiological role in lipid metabolism and are implicated in the detoxification of organophosphate (OP) insecticides. Despite the importance of OPs to agriculture and the spread of insect-borne diseases, the molecular basis for the ability of α-carboxylesterases to confer OP resistance to insects is poorly understood. In this work, we used laboratory evolution to increase the thermal stability of Lc αE7, allowing its overexpression in Escherichia coli and structure determination. The crystal structure reveals a canonical α/β-hydrolase fold that is very similar to the primary target of OPs (acetylcholinesterase) and a unique N-terminal α-helix that serves as a membrane anchor. Soaking of Lc αE7 crystals in OPs led to the capture of a crystallographic snapshot of Lc αE7 in its phosphorylated state, which allowed comparison with acetylcholinesterase and rationalization of its ability to protect insects against the effects of OPs. Finally, inspection of the active site of Lc αE7 reveals an asymmetric and hydrophobic substrate binding cavity that is well-suited to fatty acid methyl esters, which are hydrolyzed by the enzyme with specificity constants (∼10 6 M −1 s −1 ) indicative of a natural substrate. |
| Databáze: | OpenAIRE |
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