Isolation and characterization of hemorrhagic, myonecrotic and edema-inducing toxins from Bothrops insularis (jararaca ilhoa) snake venom.
Bothrops insularis snake venom was fractionated by gel filtration on Sephadex G-150 followed by ion-exchange chromatography on SP-Sephadex C-25. Two active fractions were purified to homogeneity: (1) SIII-SpI, approximate mol. wt 32,000 and N-terminal amino acid residue Val. This fraction showed esterase activity on TAME, edema-inducing activity on the rat hind paw and contractile activity on the isolated guinea pig ileum. The latter two activities were antagonized by benadryl plus methysergide; (2) SIII-SpVI, a myonecrotic and edema-inducing phospholipase, approximate mol. wt 29,000, N-terminal amino acid residue pyro-Glu, consisting of two chains of approximately 15,000 mol. wt each linked by disulphide bridge(s). The induction of edema by this fraction was not antagonized by benadryl plus methysergide, indomethacin, BW755C or BN52021, but it was antagonized by dexamethasone. Three highly purified hemorrhagic heterodimeric fractions, SIII-SpIII-3, SIII-SpIII-4 and SIII-SpIII-5, of approximate mol. wts 26,000, 29,000 and 26,000, and having N-terminal residues of Asx, Asx and Gly, respectively, were further isolated by preparative polyacrylamide slab gel electrophoresis. SIII-SpIII-4 and SIII-SpIII-5 increased the recalcification time of citrated rat plasma. None of the five isolated fractions showed any proteolytic (on casein) or kininogenase activity.
Selistre
H S
HS
Departamento de Bioquímica, Faculdade de Medicina de Riberão Preto, Universidade de São Paulo, Brasil.
Queiroz
L S
LS
Cunha
O A
OA
De Souza
G E
GE
Giglio
J R
JR
D000818:Animals; D003435:Crotalid Venoms / Q000032:analysis*; D004487:Edema / Q000139:chemically induced*; D006168:Guinea Pigs; D006470:Hemorrhage / Q000139:chemically induced*; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008970:Molecular Weight; D009132:Muscles / Q000187:drug effects* / Q000473:pathology; D009336:Necrosis; D051381:Rats; D011919:Rats, Inbred Strains; D014118:Toxins, Biological / Q000302:isolation & purification*
D003435:Crotalid Venoms; D014118:Toxins, Biological