Identification of hyaluronidase and phospholipase B in Lachesis muta rhombeata venom.
Hyaluronidases contribute to local and systemic damages after envenoming, since they act as spreading factors cleaving the hyaluronan presents in the connective tissues of the victim, facilitating the diffusion of venom components. Although hyaluronidases are ubiquitous in snake venoms, they still have not been detected in transcriptomic analysis of the Lachesis venom gland and neither in the proteome of its venom performed previously. This work purified a hyaluronidase from Lachesis muta rhombeata venom whose molecular mass was estimated by SDS-PAGE to be 60 kDa. The hyaluronidase was more active at pH 6 and 37 °C when salt concentration was kept constant and more active in the presence of 0.15 M monovalent ions when the pH was kept at 6. Venom was fractionated by reversed-phase liquid chromatography (RPLC). Edman sequencing after RPLC failed to detect hyaluronidase, but identified a new serine proteinase isoform. The hyaluronidase was identified by mass spectrometry analysis of the protein bands in SDS-PAGE. Additionally, phospholipase B was identified for the first time in Lachesis genus venom. The discovery of new bioactive molecules might contribute to the design of novel drugs and biotechnology products as well as to development of more effective treatments against the envenoming.
Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil. Electronic address: gisele.wiezel@gmail.com.
Gisele A
GA
Wiezel
Department of Physiological Sciences, Federal University of São Carlos, Rod. Washington Luís, km 235, 13565-905, São Carlos, SP, Brazil. Electronic address: patty_karina@outlook.com.
Patty K
PK
dos Santos
Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil. Electronic address: fran_acordeiro@hotmail.com.
Francielle A
FA
Cordeiro
Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil. Electronic address: karla@fcfrp.usp.br.
Karla C F
KC
Bordon
Department of Physiological Sciences, Federal University of São Carlos, Rod. Washington Luís, km 235, 13565-905, São Carlos, SP, Brazil. Electronic address: hsaraujo@ufscar.br.
Heloisa S
HS
Selistre-de-Araújo
Proteomics Resource Center, Langone Medical Center, New York University, 430 East 29th St., 8th Floor, 10016, New York, NY, USA. Electronic address: Beatrix.Ueberheide@nyumc.org.
Beatrix
B
Ueberheide
Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil. Electronic address: ecabraga@fcfrp.usp.br.
Eliane C
EC
Arantes
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D006821:Hyaluronoglucosaminidase / Q000737:chemistry* / Q000302:isolation & purification; D008245:Lysophospholipase / Q000737:chemistry* / Q000302:isolation & purification; D030162:Reptilian Proteins / Q000737:chemistry* / Q000302:isolation & purification; D014757:Viper Venoms / Q000737:chemistry / Q000201:enzymology*; D017819:Viperidae
D030162:Reptilian Proteins; D014757:Viper Venoms; D008245:Lysophospholipase; D006821:Hyaluronoglucosaminidase
Bushmaster; Hyaluronidase; Phospholipase B; Snake venom; Spreading factor; serine proteinase