Purification and characterization of patagonfibrase, a metalloproteinase showing alpha-fibrinogenolytic and hemorrhagic activities, from Philodryas patagoniensis snake venom.
Venoms of Colubridae snakes are a rich source of novel compounds, which may have applications in medicine and biochemistry. In the present study, we describe the purification and characterization of a metalloproteinase (patagonfibrase), the first protein to be isolated from Philodryas patagoniensis (Colubridae) snake venom. Patagonfibrase is a single-chain protein, showing a molecular mass of 53,224 Da and an acidic isoelectric point (5.8). It hydrolyzed selectively the Aalpha-chain of fibrinogen and when incubated with fibrinogen or plasma, the thrombin clotting time was prolonged. Prominent hemorrhage developed in mouse skin after intradermal injection of patagonfibrase. When administered into mouse gastrocnemius muscle, it induced local hemorrhage and necrosis, and systemic bleeding in lungs. Patagonfibrase showed proteolytic activity toward azocasein, which was enhanced by Ca(2+) and inhibited by Zn(2+), cysteine, dithiothreitol and Na(2)EDTA. Patagonfibrase impaired platelet aggregation induced by collagen and ADP. Thus, patagonfibrase may play a key role in the pathogenesis of disturbances that occur in P. patagoniensis envenomation, and may be used as a biological tool to explore many facets of hemostasis.
Peichoto
M E
ME
Facultad de Ciencias Exactas y Naturales y Agrimensura, Universidad Nacional del Nordeste, Corrientes, Argentina. mepeichoto@exa.unne.edu.ar
Teibler
P
P
Mackessy
S P
SP
Leiva
L
L
Acosta
O
O
Gonçalves
L R C
LR
Tanaka-Azevedo
A M
AM
Santoro
M L
ML
D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects*; D001792:Blood Platelets / Q000187:drug effects; D017812:Colubridae; D004487:Edema / Q000139:chemically induced / Q000473:pathology; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000187:drug effects* / Q000378:metabolism; D006470:Hemorrhage / Q000139:chemically induced; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007249:Inflammation / Q000139:chemically induced / Q000473:pathology; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D045726:Metalloproteases / Q000302:isolation & purification* / Q000633:toxicity*; D051379:Mice; D018482:Muscle, Skeletal / Q000187:drug effects; D009336:Necrosis / Q000139:chemically induced / Q000473:pathology; D010974:Platelet Aggregation / Q000187:drug effects; D012910:Snake Venoms / Q000201:enzymology / Q000633:toxicity*; D013696:Temperature; D013917:Thrombin / Q000187:drug effects / Q000378:metabolism; D013918:Thrombin Time
D012910:Snake Venoms; D005340:Fibrinogen; D045726:Metalloproteases; C519455:patagonfibrase, Philodryas patagoniensis; D013917:Thrombin