Y. Komori et al., BIOCHEMICAL-CHARACTERIZATION OF HEMORRHAGIC TOXIN FROM CROTALUS-VIRIDIS-VIRIDIS (PRAIRIE RATTLESNAKE) VENOM, International Journal of Biochemistry, 26(12), 1994, pp. 1411-1418
Hemorrhage, necrosis and edema are some of the effects often observed
following snake bites. This paper reports studies on the isolation and
biological properties of hemorrhagic toxin from Crotalus viridis viri
dis (Prairie rattlesnake) venom. A hemorrhagic toxin was isolated from
C. v. viridis venom by Sephadex G-50, DEAE-Sephacel and Q-Sepharose c
olumn chromatographies. The hemorrhagic toxin from C. v. viridis venom
was shown to be homogenous as demonstrated by a single band on polyac
rylamide gel electrophoresis and immunodiffusion. Its molecular weight
was approximately 54,000 daltons, and it contained 471 amino acid res
idues. The toxin possessed hemorrhagic activity with a minimum hemorrh
agic dose (MHD) of 0.11 mu g and hydrolytic activity on dimethylcasein
, casein, azocasein, azoalbumin, azocoll and hide powder azure. Hemorr
hagic and casein hydrolytic activities were inhibited by EDTA, o-phena
nthroline or dithiothreitol. The toxin contained 1 mole of zinc per mo
le of protein and zinc is essential for both hemorrhagic and proteolyt
ic activities. Hemorrhagic toxin possessed hydrolytic activity on the
B-chain of insulin, which cleaves His(5)-Leu(6), His(10)-Leu(11), Ala(
14)-Leu(15), Tyr(16)-Leu(17) and Phe(24)-Phe(25) bonds. This toxin als
o hydrolyzed Ace and BP chains of fibrinogen. Intramuscular injections
of hemorrhagic toxin caused an increase of creatine phosphokinase act
ivity in mice serum from 50.3 mU/ml to 1133 mU/ml. A toxin isolated fr
om C. v. viridis venom was shown to have strong hemorrhagic activity.
Partial characterization is reported for this major hemorrhagic toxin
in C. v. viridis venom.