Single-step purification of crotapotin and crotactine from Crotalus durissus terrificus venom using preparative isoeletric focusing
Braz. j. med. biol. res
;
30(1): 25-8, Jan. 1997. ilus
Artigo
em Inglês
| LILACS
| ID: lil-187329
RESUMO
We describe the isolation of crotoxin, a presynaptic B-neurotoxin, as well as its subunits B (crotactine) and A (crotapotin) from lyophilized Crotalus durissus terrificus venom by a single-step preparative isoelectric focusing procedure. From 98 mg of dried venom protein 20.1 mg of crotactine and 13.1 mg of crotapotin were recovered in the first step of focalization and 4.2 mg in a second run. These values correspond to 35.7 per cent of the total venom protein applied. Crotactine separated in the 9.3-7.0 pH range (tubes 1-6) and crotapotin in the 1.8-2.8 pH range (tubes 15-19) and both were homogeneous by SDS-PAGE and N-terminal amino acid analysls. Crotactine, a 12-kDa protein, presented hemolytic and phospholipase A2 activity. Thus, using isoelectric focusing we simultaneously purified both toxins in high yields. This method can be used as an altemative for the purification and characterization of proteins from other snake venoms under conditions in which biological activity is retained.
Texto completo:
DisponíveL
Índice:
LILACS (Américas)
Assunto principal:
Venenos de Serpentes
/
Crotalus
/
Venenos de Crotalídeos
/
Crotoxina
Limite:
Animais
Idioma:
Inglês
Revista:
Braz. j. med. biol. res
Assunto da revista:
Biologia
/
Medicina
Ano de publicação:
1997
Tipo de documento:
Artigo
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