Biochemical characterization of hemorrhagic toxins with fibrinogenase activity isolated from Crotalus ruber ruber venom.
Three hemorrhagic toxins with proteolytic activity were isolated from the venom of Crotalus ruber ruber (red rattlesnake). Molecular weights of HT-1, HT-2, and HT-3 were 60,000, 25,000, and 25,500, respectively. Although HT-3 was a basic protein, HT-1 and HT-2 were slightly acidic proteins. Total amino acid residues were 482,207, and 221 for HT-1, HT-2, and HT-3, respectively. Protease activity of all the toxins was inhibited in the presence of EDTA or o-phenanthroline, suggesting that the toxins are metalloproteins. Analyses for various metals by inductively coupled plasma-atomic emission spectrometry indicated that sodium, potassium, zinc, and calcium atoms were present in significant quantities. With all three toxins, there was roughly 1 mol of zinc to 1 mol of protein; the results for calcium were not consistent. All three hemorrhagic toxins degraded the A alpha chain of fibrinogen, while HT-1 also degraded the B beta chain. Although fibrinogen was degraded by the three toxins, no clots were observed, indicating that the proteolytic specificities of the three toxins were different from those of thrombin. The hemorrhagic toxins increased creatine kinase activity in mice serum, indicating muscle damage, which was substantiated by histological examination.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002942:Circular Dichroism; D003435:Crotalid Venoms / Q000032:analysis* / Q000378:metabolism / Q000633:toxicity; D005341:Fibrinolysin / Q000378:metabolism; D006470:Hemorrhage / Q000139:chemically induced; D006358:Hot Temperature; D007526:Isoelectric Point; D008970:Molecular Weight; D011487:Protein Conformation; D013379:Substrate Specificity; D013917:Thrombin / Q000032:analysis*; D014118:Toxins, Biological / Q000032:analysis*; D014357:Trypsin / Q000378:metabolism
D000596:Amino Acids; D003435:Crotalid Venoms; D014118:Toxins, Biological; C032814:hemorrhagin components (Crotalid); D014357:Trypsin; D013917:Thrombin; D005341:Fibrinolysin