Acetylcholinesterases from leaf-cutting ant atta sexdens: Purification, characterization, and capillary reactors for on-flow assays
| Autor: | Adriana M. dos Santos, Dulce Helena Ferreira de Souza, Fernando G. de Almeida, Odair Correa Bueno, Ariele C. Moreira, Bianca Rebelo Lopes, Mariana F. Fracola, Quezia B. Cass |
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| Přispěvatelé: | Universidade Federal de São Carlos (UFSCar), Universidade de São Paulo (USP), Universidade Estadual Paulista (UNESP) |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
chemistry.chemical_classification 0303 health sciences Chromatography Article Subject biology Ion chromatography Substrate (chemistry) biology.organism_classification 01 natural sciences Biochemistry Acetylcholinesterase 010602 entomology 03 medical and health sciences Hydrolysis chemistry.chemical_compound Enzyme chemistry Atta sexdens Acetylthiocholine Ammonium Molecular Biology Research Article 030304 developmental biology |
| Zdroj: | Scopus Repositório Institucional da UNESP Universidade Estadual Paulista (UNESP) instacron:UNESP Enzyme Research |
| Popis: | Made available in DSpace on 2022-04-29T08:28:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 Acetylcholinesterase (AChE) is responsible for catalyzing the hydrolysis of the neurotransmitter acetylcholine (ACh) leading to acetate and choline (Ch) release. The inhibition of AChE produces a generalized synaptic collapse that can lead to insect death. Herein we report for the first time the isolation of two AChEs from Atta sexdens which were purified by sulphate ammonium precipitation followed by ion exchange chromatography. AsAChE-A and AsAChE-B enzymes have optimum pH of 9.5 and 9.0 and higher activities in 30/50°C and 20°C, respectively, using acetylthiocholine (ATCh) as substrate. Immobilized capillary enzyme reactors (ICERs) were obtained for both enzymes (AsAChE-A-ICER and AsAChE-B-ICER) and their activities were measured by LC-MS/MS through hydrolysis product quantification of the natural substrate ACh. The comparison of activities by LC-MS/MS of both AChEs using ACh as substrate showed that AsAChE-B (free or immobilized) had the highest affinity. The inverse result was observed when the colorimetric assay (Elman method) was used for ATCh as substrate. Moreover, by mass spectrometry and phylogenetic studies, AsAChE-A and AsAChE-B were classified as belonging to AChE-2 and AChE-1 classes, respectively. Federal University of São Carlos Department of Chemistry São Paulo University Instituto de Ciências Biomédicas (ICB) São Paulo State University Center for the Study of Social Insects São Paulo State University Center for the Study of Social Insects |
| Databáze: | OpenAIRE |
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