Isolation and characterization of a myotoxic phospholipase A2 from the venom of the arboreal snake Bothriechis (Bothrops) schlegelii from Costa Rica.
A new myotoxic phospholipase A2 was isolated from the venom of the arboreal snake Bothriechis schlegelii (formerly Bothrops schlegelii) from Costa Rica, by ion-exchange chromatography on CM-Sephadex. B. schlegelii myotoxin I is a basic protein (pI > 9.3) with a subunit molecular weight of 15 kDa, which migrates as a dimer in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under nonreducing conditions. This myotoxin is recognized by antibodies generated against Bothrops asper myotoxin II (a lysine-49 phospholipase A2), by both enzyme-immunoassay and gel immunodiffusion, in the latter case with a pattern of partial identity. The toxin induces rapid myonecrosis upon intramuscular injection in mice, as evidenced by the early increase in plasma creatine kinase activity and by direct intravital microscopic observation. B. schlegelii myotoxin I also induces edema in the mouse footpad assay and exerts lethal activity (LD50 approximately 2.5 microg/g) upon intravenous injection. The toxin has a low phospholipase A2 activity (4.2 microEq.mg-1.min-1) using egg yolk phospholipids as substrate. It also shows a weak anticoagulant effect in vitro. Its N-terminal sequence, SMYELGKMILLETGKNAATSYIAYG, shows 93% homology with both Bothrops asper myotoxin II and B. jararacussu bothropstoxin I, suggesting that B. schlegelii myotoxin I may be a new lysine-49 variant of this family of myotoxic phospholipases A2.
Angulo
Y
Y
Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Chaves
E
E
Alape
A
A
Rucavado
A
A
Gutiérrez
J M
JM
Lomonte
B
B
D000818:Animals; D001483:Base Sequence; D017837:Bothrops; D003364:Costa Rica; D003435:Crotalid Venoms / Q000201:enzymology*; D054501:Group II Phospholipases A2; D051379:Mice; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D010741:Phospholipases A / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D054467:Phospholipases A2; D030162:Reptilian Proteins; D016415:Sequence Alignment
D003435:Crotalid Venoms; D009498:Neurotoxins; D030162:Reptilian Proteins; D010741:Phospholipases A; D054501:Group II Phospholipases A2; D054467:Phospholipases A2; C076780:myotoxin I