title: Revealing a Novel Otubain-Like Enzyme fromLeishmania infantumwith Deubiquitinating Activity toward K48-Linked Substrate.
abstract: Deubiquitinating enzymes (DUBs) play an important role in regulating a variety of eukaryotic processes. In this context, exploring the role of deubiquitination inLeishmania infantumcould be a promising alternative to search new therapeutic targets for leishmaniasis. Here we present the first characterization of a DUB fromL. infantum, otubain (OtuLi), and its localization within parasite. The recombinant OtuLi (rOtuLi) showed improved activity on lysine 48 (K48)-linked over K63-linked tetra-ubiquitin (Ub) and site-directed mutations on amino acids close to the catalytic site (F82) or involved in Ub interaction (L265 and F182) caused structural changes as shown by molecular dynamics, resulting in a reduction or loss of enzyme activity, respectively. Furthermore, rOtuLi stimulates lipid droplet biogenesis (an inflammatory marker) and induces IL-6 and TNF-α secretion in peritoneal macrophages, both proinflammatory cytokines. Our findings suggest that OtuLi is a cytoplasmic enzyme with K48-linked substrate specificity that could play a part in proinflammatory response in stimulated murine macrophages.
authors.affiliation: Pathogen-Host Interface Laboratory, Department of Cell Biology, University of Brasilia Brasilia, Brazil.
authors.forename: Clênia S
authors.initials: CS
authors.lastname: Azevedo
authors.affiliation: Laboratory of Electron Microscopy, Department of Cell Biology, University of Brasilia Brasilia, Brazil.
authors.forename: Bruna C
authors.initials: BC
authors.lastname: Guido
authors.affiliation: Pathogen-Host Interface Laboratory, Department of Cell Biology, University of Brasilia Brasilia, Brazil.
authors.forename: Jhonata L
authors.initials: JL
authors.lastname: Pereira
authors.affiliation: Physics Course, Catholic University of BrasiliaBrasilia, Brazil; Programa de Pós-Graduação em Ciências Genômicas e Biotecnologia, Catholic University of BrasiliaBrasilia, Brazil.
authors.forename: Diego O
authors.initials: DO
authors.lastname: Nolasco
authors.affiliation: Laboratory of Immunology and Inflammation, Department of Cell Biology, University of Brasilia Brasilia, Brazil.
authors.forename: Rafael
authors.initials: R
authors.lastname: Corrêa
authors.affiliation: Laboratory of Immunology and Inflammation, Department of Cell Biology, University of Brasilia Brasilia, Brazil.
authors.forename: Kelly G
authors.initials: KG
authors.lastname: Magalhães
authors.affiliation: Pathogen-Host Interface Laboratory, Department of Cell Biology, University of BrasiliaBrasilia, Brazil; Faculty of Ceilandia, University of BrasíliaBrasilia, Brazil.
authors.forename: Flávia N
authors.initials: FN
authors.lastname: Motta
authors.affiliation: Pathogen-Host Interface Laboratory, Department of Cell Biology, University of Brasilia Brasilia, Brazil.
authors.forename: Jaime M
authors.initials: JM
authors.lastname: Santana
authors.affiliation: UMR 7245 Centre Nationnal de la Recherche Scientifique, Muséum National d'Histoire Naturelle, Sorbonne Universités Paris, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Grellier
authors.affiliation: Pathogen-Host Interface Laboratory, Department of Cell Biology, University of Brasilia Brasilia, Brazil.
authors.forename: Izabela M D
authors.initials: IM
authors.lastname: Bastos
keywords: cysteine protease; deubiquitination; leishmania; molecular dynamic; otubain; site-directed mutagenesis