title: BJ-PI2, a non-hemorrhagic metalloproteinase from Bothrops jararaca snake venom.
abstract: BACKGROUND
  Envenoming by Bothrops jararaca can result in local pain, edema, hemorrhage and necrosis, partially mediated by snake venom metalloproteinases (SVMPs). Here, we describe the characterization of BJ-PI2, a P-I class SVMP from B. jararaca venom, and its local tissue actions.
  
  
  METHODS
  BJ-PI2 was purified by a combination of gel filtration, anion-exchange chromatography and reverse phase HPLC, and identified by mass spectrometry. Clotting and fibrin(ogen)olytic activities were assayed using conventional methods. Hemorrhagic activity and changes in vascular permeability were examined in rat dorsal skin. Myonecrosis and inflammatory activity were examined in mouse gastrocnemius muscle.
  
  
  RESULTS
  BJ-PI2 was a 23.08kDa single-chain polypeptide. Tryptic fragments showed highest homology with SVMP insularinase A from Bothrops insularis, but also with B. jararaca SVMP bothrojaractivase; less similarity was observed with B. jararaca SVMPs BJ-PI and jararafibrases II and IV. BJ-PI2 did not clot fibrinogen or rat citrated plasma but had α- and β-fibrinogenolytic activity (inhibited by EDTA and 1,10-phenanthroline but not by PMSF) and attenuated coagulation after plasma recalcification. BJ-PI2 had fibrinolytic activity. BJ-PI2 increased the vascular permeability of rat dorsal skin (inhibited by 1,10-phenanthroline). BJ-PI2 was not hemorrhagic or myonecrotic but caused migration of inflammatory cells. In contrast, venom was strongly hemorrhagic and myonecrotic but caused less infiltration of inflammatory cells.
  
  
  CONCLUSIONS
  BJ-PI2 is a non-hemorrhagic, non-myonecrotic, non-coagulant P-I class SVMP that may enhance vascular permeability and inflammatory cell migration in vivo.
  
  
  CONCLUSIONS
  BJ-PI2 contributes to enhanced vascular permeability and inflammatory cell migration after envenoming, but not to venom-induced hemorrhage and necrosis.
authors.affiliation: Departamento de Farmacologia, Universidade Estadual de Campinas, Campinas, Brazil.
authors.forename: Igor Rapp Ferreira
authors.initials: IR
authors.lastname: da Silva
authors.forename: Raquel
authors.initials: R
authors.lastname: Lorenzetti
authors.forename: André Lisboa
authors.initials: AL
authors.lastname: Rennó
authors.forename: Lineu
authors.initials: L
authors.lastname: Baldissera
authors.forename: André
authors.initials: A
authors.lastname: Zelanis
authors.forename: Solange Maria de Toledo
authors.initials: SM
authors.lastname: Serrano
authors.forename: Stephen
authors.initials: S
authors.lastname: Hyslop
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops; D002199:Capillary Permeability / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000302:isolation & purification / Q000494:pharmacology; D006470:Hemorrhage / Q000139:chemically induced; D013058:Mass Spectrometry; D045726:Metalloproteases / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats
chemical_list: D003435:Crotalid Venoms; C576279:BJ-PI2 non-hemorrhagic metalloproteinase, Bothrops jararaca; D045726:Metalloproteases