Isolation and characterization of a hemorrhagic proteinase from timber rattlesnake venom

Abstract

A protein isolated from timber rattlesnake (Crotalus horridus horridus) venom by ion-exchange and high-pressure liquid chromatography is hemorrhage inducing and lethal to mice (LD50 of 10 micrograms/g of body weight). It is a Ca2+- and Zn2+-containing proteinase and has the ability to hydrolyze hide powder azure. Atomic absorption spectroscopy shows 2.5 Ca2+ and 1 Zn2+ per protein monomer. The proteinase activity is destroyed by incubation with disulfide-reducing agents and by dialysis against ethylenediaminetetraacetate. Coincident with the loss of proteinase activity is a corresponding loss of lethal and hemorrhagic activities, suggesting that all three are related. Attempts to replace the metals and restore activity have been unsuccessful. Amino acid analysis and isoelectric focusing reveal that this component is an acidic protein (pI = 5.1) containing about 20 disulfide bonds and 507 residues. Reduction of one disulfide bond per molecule decreases proteinase activity by 50% while reduction of eight disulfide bonds decreases activity by 80%. Loss of hemorrhagic activity parallels the decrease in proteinase activity.

‘Partitagin’ a hemorrhagic metalloprotease from Hippasa partita spider venom: Role in tissue necrosis

“…However, its role in maintaining the structural integrity cannot be ignored. The metal content appears to be bit more than that reported for several SVMPs, however, agrees well with the metal content of Lachesis muta muta [27], Agkistrodon acutus [28], Crotalus horridus horridus [29].…”

Characterization of inhibitor A, a protease from Bacillus thuringiensis which degrades attacins and cecropins, two classes of antibacterial proteins in insects

“…The mechanism for this has not been established but the efficient hydrolysis of the collagen-containing substrate Hide Powder may indicate that InA has an hemorrhagic activity. A toxic metalloprotease with such properties has recently been isolated from rattlesnake venom [27]. The final assessment of the physiological role of InA in the course of an infection of an insect must therefore await further studies in vivo where survival of both host and parasite as well as InA, attacin and cecropin concentrations are recorded simultaneously.…”

Inventory of α- and β-Fibrinogenases from Snake Venoms

“…Hemorrhagic proteinase HPIV from Crotalus h. horridus venom, has a molecular weight of 57,ff[ and a pI of 5.1. This protein is a zinc metalloproteinase possessing Ao,->BB-chain degrading activity with fibrinogen as substrate, and fibrin o-chain degrading activity (77,78).…”