Purification and partial characterization of the hemorrhagic factor from the venom of Crotalus adamanteus (eastern diamondback rattlesnake).
A proteinase from Crotalus adamanteus venom has been isolated to the stage of electrophoretic homogeneity by chromatography of the crude venom on CM-Sepharose, Phenyl-Sepharose, Ultrogel AcA 44 and DEAE-Sepharose. The proteinase accounts for 100% of the detectable hemorrhagic activity in the venom, and the name C. adamanteus proteinase H is suggested. Proteinase H is a single chain glycoprotein with a molecular weight of 85,700. Proteinase H is active on casein and hide powder azure, but does not digest benzoyl-L-arginine ethyl ester or benzoyl-L-tyrosine ethyl ester. Proteolytic and hemorrhagic activity were both abolished by treatment with EDTA and neither activity was restored by prolonged dialysis against Zn2+ or Ca2+. However, both activities were retained after treatment with phenylmethylsulfonyl fluoride. A minimal hemorrhagic response was elicited in mice by s.c. injection of 0.1 microgram of proteinase H. The caseinolytic activity of proteinase H was not inhibited during incubation with human alpha 2-macroglobulin, nor was the inhibitor inactivated by proteinase H.
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D000818:Animals; D002364:Caseins / Q000378:metabolism; D003435:Crotalid Venoms / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000302:isolation & purification* / Q000494:pharmacology; D006470:Hemorrhage / Q000139:chemically induced*; D006801:Humans; D008666:Metalloendopeptidases; D008970:Molecular Weight; D000511:alpha-Macroglobulins / Q000494:pharmacology
D002364:Caseins; D003435:Crotalid Venoms; D000511:alpha-Macroglobulins; D010450:Endopeptidases; C046858:Crotalus adamanteus proteinase H; D008666:Metalloendopeptidases