Purification and partial characterization of the hemorrhagic factor from the venom of Crotalus adamanteus (eastern diamondback rattlesnake)

• Published in: Toxicon (Oxford) Vol. 23; no. 4; p. 657
• Main Authors: Kurecki, T, Kress, L F
• Format: Journal Article
• Language: English
• Published: England 1985
• Subjects: alpha-Macroglobulins - pharmacology; Animals; Caseins - metabolism; Crotalid Venoms - analysis; Electrophoresis, Polyacrylamide Gel; Endopeptidases - isolation & purification; Endopeptidases - pharmacology; Hemorrhage - chemically induced; Humans; Metalloendopeptidases; Molecular Weight
• ISSN: 0041-0101; 1879-3150
• DOI: 10.1016/0041-0101(85)90370-8

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.