Structural features and development of an assay platform of the parasite target deoxyhypusine synthase of brugia malayi and leishmania major

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dc.contributor.authorSilva, Suélen Fernandes [UNESP]
dc.contributor.authorKlippel, Angélica Hollunder [UNESP]
dc.contributor.authorRamos, Priscila Zonzini
dc.contributor.authorSantiago, André da Silva
dc.contributor.authorValentini, Sandro Roberto [UNESP]
dc.contributor.authorBengtson, Mario Henrique
dc.contributor.authorMassirer, Katlin Brauer
dc.contributor.authorBilsland, Elizabeth
dc.contributor.authorCouñago, Rafael Miguez
dc.contributor.authorZanelli, Cleslei Fernando [UNESP]
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionUniversidade Estadual de Campinas (UNICAMP)
dc.date.accessioned2021-06-25T11:06:11Z
dc.date.available2021-06-25T11:06:11Z
dc.date.issued2020-10-01
dc.description.abstractDeoxyhypusine synthase (DHS) catalyzes the first step of the post-translational modification of eukaryotic translation factor 5A (eIF5A), which is the only known protein containing the amino acid hypusine. Both proteins are essential for eukaryotic cell viability, and DHS has been suggested as a good candidate target for small molecule-based therapies against eukaryotic pathogens. In this work, we focused on the DHS enzymes from Brugia malayi and Leishmania major, the causative agents of lymphatic filariasis and cutaneous leishmani-asis, respectively. To enable B. malayi (Bm)DHS for future target-based drug discovery pro-grams, we determined its crystal structure bound to cofactor NAD+. We also reported an in vitro biochemical assay for this enzyme that is amenable to a high-throughput screening for-mat. The L. major genome encodes two DHS paralogs, and attempts to produce them recombinantly in bacterial cells were not successful. Nevertheless, we showed that ectopic expression of both LmDHS paralogs can rescue yeast cells lacking the endogenous DHS-encoding gene (dys1). Thus, functionally complemented dys1Δ yeast mutants can be used to screen for new inhibitors of the L. major enzyme. We used the known human DHS inhibi-tor GC7 to validate both in vitro and yeast-based DHS assays. Our results show that BmDHS is a homotetrameric enzyme that shares many features with its human homologue, whereas LmDHS paralogs are likely to form a heterotetrameric complex and have a distinct regulatory mechanism. We expect our work to facilitate the identification and development of new DHS inhibitors that can be used to validate these enzymes as vulnerable targets for therapeutic interventions against B. malayi and L. major infections.en
dc.description.affiliationChemistry Institute São Paulo State University—UNESP
dc.description.affiliationSchool of Pharmaceutical Sciences São Paulo State University—UNESP
dc.description.affiliationMolecular Biology and Genetic Engineering Center (CBMEG) Medicinal Chemistry Center (CQMED) Structural Genomics Consortium (SGC-UNICAMP) University of Campinas-UNICAMP
dc.description.affiliationDepartment of Structural and Functional Biology Institute of Biology University of Campinas—UNICAMP
dc.description.affiliationDepartment of Biochemistry and Tissue Biology Institute of Biology University of Campinas— UNICAMP
dc.description.affiliationUnespChemistry Institute São Paulo State University—UNESP
dc.description.affiliationUnespSchool of Pharmaceutical Sciences São Paulo State University—UNESP
dc.format.extent1-31
dc.identifierhttp://dx.doi.org/10.1371/journal.pntd.0008762
dc.identifier.citationPLoS Neglected Tropical Diseases, v. 14, n. 10, p. 1-31, 2020.
dc.identifier.doi10.1371/journal.pntd.0008762
dc.identifier.issn1935-2735
dc.identifier.issn1935-2727
dc.identifier.lattes1525665408900195
dc.identifier.orcid0000-0001-7831-1149
dc.identifier.scopus2-s2.0-85094219500
dc.identifier.urihttp://hdl.handle.net/11449/208090
dc.language.isoeng
dc.relation.ispartofPLoS Neglected Tropical Diseases
dc.sourceScopus
dc.titleStructural features and development of an assay platform of the parasite target deoxyhypusine synthase of brugia malayi and leishmania majoren
dc.typeArtigo
unesp.author.lattes1525665408900195[10]
unesp.author.orcid0000-0001-7831-1149[10]

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