Identification and characterization of a recombinant cysteine peptidase (AsCathL) from leaf-cutting ant Atta sexdens Linnaeus, 1758 (Hymenoptera, Formicidae)

dc.contributor.authorSantos Correa, Katia Celina
dc.contributor.authorMoreira, Ariele Cristina
dc.contributor.authorAbd El-Raheem Ibrahim, Amr Galal
dc.contributor.authorRamos de Jesus, Hugo César
dc.contributor.authorMicocci, Kelli Cristina [UNESP]
dc.contributor.authorCrizóstomo Kock, Flávio Vinícius
dc.contributor.authorBueno, Odair C. [UNESP]
dc.contributor.authorVenâncio, Tiago
dc.contributor.authorHenrique-Silva, Flávio
dc.contributor.authorSouza, Dulce Helena F.
dc.contributor.institutionUniversidade Federal de São Carlos (UFSCar)
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.contributor.institutionUniversidade Estadual Paulista (UNESP)
dc.date.accessioned2023-07-29T13:21:22Z
dc.date.available2023-07-29T13:21:22Z
dc.date.issued2023-01-01
dc.description.abstractCysteine peptidases are involved in physiological processes of insect development and have been considered as potential targets for the development of insect control strategies. In this study, we obtained a recombinant cysteine cathepsin L (AsCathL) from leaf-cutting ant (Atta sexdens), a species from the order Hymenoptera who causes enormous damage to crops, natural forests and reforested areas. RT-qPCR showed AsCathL expression throughout insect development and in all body parts of the adult insect analysed, suggesting its role as a lysosomal cathepsin. AsCathL encodes a protein of 320 amino acid residues consisting of a pro-peptide and the mature with amino acids sequence over 67% similarity with lysosomal cathepsin L of species from Lepidoptera and Diptera. Phylogenetic tree revealed that AsCathL is very similar to predicted cathepsins found in other ants. Recombinant AsCathL was expressed in insoluble form by Escherichia coli Arctic Express (DE3) RIL, purified under denaturing conditions and refolded. The enzyme showed hydrolytic activity in vitro towards synthetic substrate Z-Phe-Arg-AMC at acidic pH. Synthetic inhibitor E−64 acted against peptidase activity and a study regarding the interaction between E−64 and AsCathL using nuclear magnetic resonance (NMR) revealed that 83.18% from all E−64 molecules are irreversibly bound to AsCathL. In addition, the proteolytic activity of AsCathL was strongly inhibited by recombinant sugarcane cystatins with Ki ranging from 0.6 nM to 2.95 nM. To the best of our knowledge this is the first report characterizing a cysteine peptidase from leaf-cutting ants, which may contribute to future studies of ants’ cathepsins.en
dc.description.affiliationDepartment of Chemistry Federal University of São Carlos
dc.description.affiliationDepartment of Computing and Mathematics at University of São Paulo, Ribeirão Preto, SP
dc.description.affiliationCenter for the Study of Social Insects UNESP - São Paulo State University, SP
dc.description.affiliationDepartment of Genetics and Evolution Federal University of São Carlos
dc.description.affiliationUnespCenter for the Study of Social Insects UNESP - São Paulo State University, SP
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipIdCNPq: 141424/2020–6
dc.description.sponsorshipIdFAPESP: 2014/12169–2
dc.description.sponsorshipIdFAPESP: 2015/21517–7
dc.description.sponsorshipIdFAPESP: 2017/15455–4
dc.description.sponsorshipIdFAPESP: 2018/06297–9
dc.description.sponsorshipIdFAPESP: 2018/16040–5
dc.identifierhttp://dx.doi.org/10.1016/j.pep.2022.106174
dc.identifier.citationProtein Expression and Purification, v. 201.
dc.identifier.doi10.1016/j.pep.2022.106174
dc.identifier.issn1096-0279
dc.identifier.issn1046-5928
dc.identifier.scopus2-s2.0-85138164598
dc.identifier.urihttp://hdl.handle.net/11449/247627
dc.language.isoeng
dc.relation.ispartofProtein Expression and Purification
dc.sourceScopus
dc.subjectAtta sexdens
dc.subjectCysteine cathepsin
dc.subjectCysteine peptidase
dc.subjectInsect cathepsin
dc.subjectSugarcane cystatin
dc.titleIdentification and characterization of a recombinant cysteine peptidase (AsCathL) from leaf-cutting ant Atta sexdens Linnaeus, 1758 (Hymenoptera, Formicidae)en
dc.typeArtigo
unesp.author.orcid0000-0001-8479-6744[10]

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