title: Biochemical characterization and pharmacological properties of new basic PLA2 BrTX-I isolated from Bothrops roedingeri (Roedinger's Lancehead) Mertens, 1942, snake venom.
abstract: BrTX-I, a PLA2, was purified from Bothrops roedingeri venom after only one chromatographic step using reverse-phase HPLC on μ-Bondapak C-18 column. A molecular mass of 14358.69 Da was determined by MALDI-TOF mass spectrometry. Amino acid analysis showed a high content of hydrophobic and basic amino acids as well as 14 half-cysteine residues. The total amino acid sequence was obtained using SwissProt database and showed high amino acid sequence identity with other PLA2 from snake venom. The amino acid composition showed that BrTX-I has a high content of Lys, Tyr, Gly, Pro, and 14 half-Cys residues, typical of a basic PLA2. BrTX-I presented PLA2 activity and showed a minimum sigmoidal behavior, reaching its maximal activity at pH 8.0, 35-45°C, and required Ca(2+). In vitro, the whole venom and BrTX-I caused a neuromuscular blockade in biventer cervicis preparations in a similar way to other Bothrops species. BrTX-I induced myonecrosis and oedema-forming activity analyzed through injection of the purified BrTX-I in mice. Since BrTX-I exerts a strong proinflammatory effect, the enzymatic phospholipid hydrolysis might be relevant for these phenomena; incrementing levels of IL-1, IL-6, and TNF α were observed at 15 min, 30 min, one, two, and six hours postinjection, respectively.
authors.affiliation: Department of Biochemistry, Institute of Biology, State University of Campinas (UNICAMP), P.O. Box 6109, 13083-970 Campinas, SP, Brazil.
authors.forename: Mauricio Aurelio
authors.initials: MA
authors.lastname: Gomes Heleno
authors.forename: Paulo Aparecido
authors.initials: PA
authors.lastname: Baldasso
authors.forename: Luis Alberto
authors.initials: LA
authors.lastname: Ponce-Soto
authors.forename: Sérgio
authors.initials: S
authors.lastname: Marangoni
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D017837:Bothrops; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000737:chemistry; D016207:Cytokines / Q000378:metabolism; D004487:Edema / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007249:Inflammation; D008297:Male; D051379:Mice; D010455:Peptides / Q000737:chemistry; D054467:Phospholipases A2 / Q000737:chemistry* / Q000494:pharmacology; D054497:Phospholipases A2, Secretory / Q000737:chemistry* / Q000494:pharmacology; D030162:Reptilian Proteins / Q000737:chemistry* / Q000494:pharmacology; D012910:Snake Venoms / Q000201:enzymology*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013696:Temperature
chemical_list: D000596:Amino Acids; D016207:Cytokines; D010455:Peptides; D030162:Reptilian Proteins; D012910:Snake Venoms; C584303:BrTx-I protein, Bothrops roedingeri; D054467:Phospholipases A2; D054497:Phospholipases A2, Secretory; D003545:Cysteine