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Purification and biochemical characterization of TsMS 3 and TsMS 4: neuropeptide-degrading metallopeptidases in the tityus serrulatus venom
Cajado-Carvalho, Daniela; Silva, Cristiane Castilho Fernandes da; Kodama, Roberto Tadashi; Mariano, Douglas Oscar Ceolin; Pimenta, Daniel Carvalho; Duzzi, Bruno; Kuniyoshi, Alexandre Kazuo; Portaro, Fernanda Calheta Vieira.
  • Cajado-Carvalho, Daniela; Instituto Butantan. Laboratório de Imunoquímica.
  • Silva, Cristiane Castilho Fernandes da; Instituto Butantan. Laboratório de Imunoquímica.
  • Kodama, Roberto Tadashi; Instituto Butantan. Laboratório de Imunoquímica.
  • Mariano, Douglas Oscar Ceolin; Instituto Butantan. Laboratório de Bioquímica e Biofísica.
  • Pimenta, Daniel Carvalho; Instituto Butantan. Laboratório de Bioquímica e Biofísica.
  • Duzzi, Bruno; Instituto Butantan. Laboratório de Imunoquímica.
  • Kuniyoshi, Alexandre Kazuo; Instituto Butantan. Laboratório de Imunoquímica.
  • Portaro, Fernanda Calheta Vieira; Instituto Butantan. Laboratório de Imunoquímica.
Toxins ; 11(4): 194, 2019.
Article en En | SES-SP, SESSP-IBPROD, SES-SP | ID: but-ib15951
Biblioteca responsable: BR78.1
Ubicación: BR78.1
ABSTRACT
Although omics studies have indicated presence of proteases on the Tityus serrulatus venom (TsV), little is known about the function of these molecules. The TsV contains metalloproteases that cleave a series of human neuropeptides, including the dynorphin A (1-13) and the members of neuropeptide Y family. Aiming to isolate the proteases responsible for this activity, the metalloserrulase 3 and 4 (TsMS 3 and TsMS 4) were purified after two chromatographic steps and identified by mass spectrometry analysis. The biochemical parameters (pH, temperature and cation effects) were determined for both proteases, and the catalytic parameters (Km, kcat, cleavage sites) of TsMS 4 over fluorescent substrate were obtained. The metalloserrulases have a high preference for cleaving neuropeptides but presented different primary specificities. For example, the Leu-enkephalin released from dynorphin A (1-13) hydrolysis was exclusively performed by TsMS 3. Neutralization assays using Butantan Institute antivenoms show that both metalloserrulases were well blocked. Although TsMS 3 and TsMS 4 were previously described through cDNA library studies using the venom gland, this is the first time that both these toxins were purified. Thus, this study represents a step further in understanding the mechanism of scorpion venom metalloproteases, which may act as possible neuropeptidases in the envenomation process.
Texto completo: 1 Colección SES: Producao_cientifica Banco de datos: SES-SP / SESSP-IBPROD Idioma: En Año: 2019 Tipo del documento: Article
Texto completo: 1 Colección SES: Producao_cientifica Banco de datos: SES-SP / SESSP-IBPROD Idioma: En Año: 2019 Tipo del documento: Article