Presence of heat-stable hemorrhagic toxins in snake venoms.
Twenty-eight snake venoms (seven Agkistrodon venoms, six Bothrops venoms, 13 Crotalus venoms, one Sistrurus venom, and one Bitis venom) were examined for the presence of heat-stable (100 degrees C, 5 min) hemorrhagic toxins. Both heated and unheated venoms were analyzed for their protein composition by SDS-PAGE, and tested for their hemorrhagic activity in vivo in mice and for their proteolytic activity on two different substrates. Heating all venoms led to the denaturation and loss of some proteins; however, most of the venoms retained a significant number of proteins. Seventeen venoms contained more than seven proteins after heating, whereas five venoms contained only one to three proteins. All but nine of the heated venoms had substantial hemorrhagic activity, and Agkistrodon piscivorus piscivorus venom had very high activity, almost four times that of the second most hemorrhagic venom from Crotalus viridis lutosus. Most venoms, heated or unheated, had activity on the two protease substrates. Using succinylated casein as the substrate, there was a wide range of activity, and heating drastically reduced the activity of all venoms except that of Crotalus ruber and Crotalus molossus molossus. With azocoll as substrate, all but two of the unheated venoms (Crotalus adamanteus and Crotalus viridis concolor) had very high activity, whereas upon heating, all except five venoms lost essentially all of their activity. Heated venoms from snakes in the Agkistrodon genus (except for Agkistrodon blomhoffi blomhoffi, an Asian snake) retained activity on azocoll, and this activity tended to correlate better with hemorrhagic activity of the venom than did proteolytic activity on casein.
Ownby
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Department of Physiological Sciences, Oklahoma State University, Stillwater 74078.
Colberg
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D000818:Animals; D001391:Azo Compounds / Q000378:metabolism; D002364:Caseins / Q000378:metabolism; D003094:Collagen / Q000378:metabolism; D004396:Coloring Agents / Q000378:metabolism; D003435:Crotalid Venoms / Q000378:metabolism / Q000633:toxicity; D004591:Electrophoresis, Polyacrylamide Gel; D006470:Hemorrhage / Q000139:chemically induced*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D051379:Mice; D011489:Protein Denaturation; D011506:Proteins / Q000032:analysis*; D012910:Snake Venoms / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D014118:Toxins, Biological / Q000032:analysis* / Q000633:toxicity
D001391:Azo Compounds; D002364:Caseins; D004396:Coloring Agents; D003435:Crotalid Venoms; D011506:Proteins; D012910:Snake Venoms; D014118:Toxins, Biological; C028470:Azocoll; D003094:Collagen