Biochemical characterization and assessment of leishmanicidal effects of a new L-amino acid oxidase from Crotalus durissus collilineatus snake venom (CollinLA AO-I).
de Freitas, Vitor; Costa, Tássia Rafaella; Nogueira, Amanda Rodrigueset al.
[en] This study reports the isolation of CollinLAAO-I, a new L-amino acid oxidase from Crotalus durissus collilineatus snake venom, its biochemical characterization and leishmanicidal potential in Leishmania spp. CollinLAAO-I (63.1 kDa) was successfully isolated with high purity using two chromatographic steps and represents 2.5% of total venom proteins. CollinLAAO-I displayed high enzymatic activity (4262.83 U/mg/min), significantly reducing after 28 days. The enzymatic activity of CollinLAAO-I revealed higher affinity for hydrophobic amino acids such as L-leucine, high enzymatic activity in a wide pH range (6.0-10.0), at temperatures from 0 to 25 °C, and showed complete inhibition in the presence of Na+ and K+. Cytotoxicity assays revealed IC50 of 18.49 and 11.66 μg/mL for Leishmania (L.) amazonensis and Leishmania (L.) infantum, respectively, and the cytotoxicity was completely suppressed by catalase. CollinLAAO-I significantly increased the intracellular concentration of reactive oxygen species (ROS) and reduced the mitochondrial potential of both Leishmania species. Furthermore, CollinLAAO-I decreased the parasite capacity to infect macrophages by around 70%, indicating that even subtoxic concentrations of CollinLAAO-I can interfere with Leishmania vital processes. Thus, the results obtained for CollinLAAO-I provide important support for developing therapeutic strategies against leishmaniasis.
Disciplines :
Chemistry
Author, co-author :
de Freitas, Vitor; Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Costa, Tássia Rafaella; Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Nogueira, Amanda Rodrigues; Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Polloni, Lorena; Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Alves de Melo Fernandes, Thales; Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Correia, Lucas Ian Veloso; Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Borges, Bruna Cristina; Laboratory of Osteoimmunology and Tumor Immunology, Institute of Biomedical Sciences, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Teixeira, Samuel Cota; Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Silva, Marcelo José Barbosa; Laboratory of Osteoimmunology and Tumor Immunology, Institute of Biomedical Sciences, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Amorim, Fernanda Gobbi; Laboratory of Mass Spectrometry, Department of Chemistry, MolSys-RU, University of Liège, Liège, Belgium
Quinton, Loïc ; Université de Liège - ULiège > Département de chimie (sciences) > Chimie biologique ; Université de Liège - ULiège > Molecular Systems (MolSys) ; Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Saraiva, André Lopes; Laboratory of Biochemistry and Molecular Biology, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Espindola, Foued Salmen; Laboratory of Biochemistry and Molecular Biology, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Iwai, Leo Kei; Laboratory of Applied Toxinology (LETA) and Center of Toxins, Immune-Response and Cell Signaling (CeTICS), Butantan Institute, São Paulo, SP, Brazil
Rodrigues, Renata Santos; Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
Yoneyama, Kelly Aparecida Geraldo; Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil
de Melo Rodrigues Ávila, Veridiana ; Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil. Electronic address: veridiana@ufu.br
Biochemical characterization and assessment of leishmanicidal effects of a new L-amino acid oxidase from Crotalus durissus collilineatus snake venom (CollinLA AO-I).
The authors thank the Federal University of Uberlândia ( UFU ), Brazil, and the Brazilian funding agencies CNPq , CAPES and FAPEMIG for providing financial support to this study: FAPEMIG ( CBB - APQ-01401-17 ) and National Institute of Sciences and Technology in Theranostics and Nanobiotechnology INCT-TeraNano- CNPq / CAPES / FAPEMIG , Grant numbers: CNPq- 465669/2014–0 , CAPES: 88887.136343/2017–00 and FAPEMIG: APQ-03613-17 .The authors thank the Federal University of Uberlândia (UFU), Brazil, and the Brazilian funding agencies CNPq, CAPES and FAPEMIG for providing financial support to this study: FAPEMIG (CBB - APQ-01401-17) and National Institute of Sciences and Technology in Theranostics and Nanobiotechnology INCT-TeraNano-CNPq/CAPES/FAPEMIG, Grant numbers: CNPq-465669/2014–0, CAPES: 88887.136343/2017–00 and FAPEMIG: APQ-03613-17.
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