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Acetylcholinesterases from Leaf-Cutting ant Atta sexdens: Purification, Characterization, and Capillary Reactors for On-Flow Assays.
Dos Santos, Adriana M; Moreira, Ariele C; Lopes, Bianca Rebelo; Fracola, Mariana F; de Almeida, Fernando G; Bueno, Odair C; Cass, Quezia B; Souza, Dulce Helena F.
Afiliação
  • Dos Santos AM; Federal University of São Carlos, Department of Chemistry, São Carlos, SP, Brazil.
  • Moreira AC; Federal University of São Carlos, Department of Chemistry, São Carlos, SP, Brazil.
  • Lopes BR; Federal University of São Carlos, Department of Chemistry, São Carlos, SP, Brazil.
  • Fracola MF; Federal University of São Carlos, Department of Chemistry, São Carlos, SP, Brazil.
  • de Almeida FG; São Paulo University, Instituto de Ciências Biomédicas (ICB), São Paulo, Brazil.
  • Bueno OC; São Paulo State University, Center for the Study of Social Insects, Rio Claro, SP, Brazil.
  • Cass QB; Federal University of São Carlos, Department of Chemistry, São Carlos, SP, Brazil.
  • Souza DHF; Federal University of São Carlos, Department of Chemistry, São Carlos, SP, Brazil.
Enzyme Res ; 2019: 6139863, 2019.
Article em En | MEDLINE | ID: mdl-31354985
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Enzyme Res Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Enzyme Res Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos