Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/23422
Title: Screening for target toxins of the antiophidic protein DM64 through a gel-based interactomics approach
Authors: Rocha, Surza L. G.
Ferreira, Ana G. C. Neves
Trugilho, Monique R. O.
Angulo, Yamileth
Lomonte, Bruno
Valente, Richard H.
Domont, Gilberto B.
Perales, Jonas
Affilliation: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Toxinologia. Rio de Janeiro, RJ. Brasil / Instituto Nacional de Ciência e Tecnologia em Toxinas, CNPq. Brasília, DF, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Toxinologia. Rio de Janeiro, RJ. Brasil / Instituto Nacional de Ciência e Tecnologia em Toxinas, CNPq. Brasília, DF, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Toxinologia. Rio de Janeiro, RJ. Brasil / Instituto Nacional de Ciência e Tecnologia em Toxinas, CNPq. Brasília, DF, Brasil.
Universidad de Costa Rica. Facultad de Microbiologia. Instituto Clodomiro Picado. San José, Costa Rica.
Universidad de Costa Rica. Facultad de Microbiologia. Instituto Clodomiro Picado. San José, Costa Rica.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Toxinologia. Rio de Janeiro, RJ. Brasil / Instituto Nacional de Ciência e Tecnologia em Toxinas, CNPq. Brasília, DF, Brasil.
Universidade Federal do Rio de Janeiro, Instituto de Química. Departamento de Bioquímica. Laboratório de Química de Proteínas/ Unidade Proteômica. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Toxinas, CNPq. Brasília, DF, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Toxinologia. Rio de Janeiro, RJ. Brasil / Instituto Nacional de Ciência e Tecnologia em Toxinas, CNPq. Brasília, DF, Brasil.
Abstract: DM64 is a glycosylated protein with antivenom activity isolated from the serum of the opossum Didelphis aurita. It binds non-covalently to myotoxins I (Asp49) and II (Lys49) from Bothrops asper venom and inhibits their myotoxic effect. In this study, an affinity column with immobilized DM64 as bait was used to fish potential target toxins. All ten isolated myotoxins tested were able to effectively bind to the DM64 column. To better access the specificity of the inhibitor, crude venoms from Bothrops (8 species), Crotalus (2 species) and Naja naja atra were submitted to the affinity purification. Venom fractions bound and nonbound to the DM64 column were analyzed by two-dimensional gel electrophoresis and MALDI-TOF/TOF MS. Although venom fractions bound to the column were mainly composed of basic PLA2, a few spots corresponding to acidic PLA2 were also observed. Some unexpected protein spots were also identified: C-type lectins and CRISP may represent putative new targets for DM64, whereas the presence of serine peptidases in the venom bound fraction is likely a consequence of nonspecific binding to the column matrix. The present results contribute to better delineate the inhibitory potential of DM64, providing a framework for the development of more specific antivenom therapies.
Keywords: Snake venoms
Myotoxin
Myotoxin inhibitor
Didelphis aurita
Proteomics
Interactomics
keywords: Venenos de Serpentes
Proteômica
Miotoxina
Inibidor de miotoxina
Didelphis aurita
Issue Date: 2017
Publisher: Elsevier
Citation: ROCHA, Surza L. G. et al. Screening for target toxins of the antiophidic protein DM64 through a gel-based interactomics approach. Journal of Proteomics, v.151, p.204-213, 2017.
DOI: 10.1016/j.jprot.2016.05.020
ISSN: 1874-3919
Copyright: restricted access
Appears in Collections:IOC - Artigos de Periódicos

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