BJ-PI2, A non-hemorrhagic metalloproteinase from Bothrops jararaca snake venom

• Published in: Biochimica et biophysica acta Vol. 1820; no. 11; pp. 1809 - 1821
• Main Authors: da Silva, Igor Rapp Ferreira, Lorenzetti, Raquel, Rennó, André Lisboa, Baldissera, Lineu, Zelanis, André, Serrano, Solange Maria de Toledo, Hyslop, Stephen
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.11.2012
• Subjects: Amino Acid Sequence; Animals; anion exchange chromatography; Bothrops; Bothrops jararaca; Capillary Permeability - drug effects; cell movement; Cell Movement - drug effects; coagulation; Coagulopathy; Crotalid Venoms - chemistry; Crotalid Venoms - enzymology; Crotalid Venoms - isolation & purification; Crotalid Venoms - pharmacology; edema; EDTA (chelating agent); fibrin; fibrinogen; gel chromatography; Hemorrhage; Hemorrhage - chemically induced; Mass Spectrometry; Metalloproteases - chemistry; Metalloproteases - isolation & purification; Metalloproteases - pharmacology; metalloproteinases; Mice; Molecular Sequence Data; muscles; Myonecrosis; necrosis; pain; permeability; polypeptides; Rats; reversed-phase high performance liquid chromatography; reversed-phase liquid chromatography; Snake venom metalloproteinase (SVMP); snakes; Vascular permeability; venoms; Myonecrosis; Snake venom metalloproteinase (SVMP); Bothrops jararaca; Hemorrhage; Vascular permeability; Coagulopathy
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2012.07.011

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. 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. 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. 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. BJ-PI2 contributes to enhanced vascular permeability and inflammatory cell migration after envenoming, but not to venom-induced hemorrhage and necrosis. ► Bothrops jararaca venom contains snake venom metalloproteinases (SVMPs). ► BJ-PI2 is a non-hemorrhagic, non-myotoxic, non-coagulant class P-I SVMP. ► BJ-PI2 increases vascular permeability and causes inflammatory cell migration. ► BJ-PI2 may contribute to venom-induced vascular permeability and inflammation.