Studies on the mechanism of hemorrhage production by five proteolytic hemorrhagic toxins from Crotalus atrox venom.
The sites and relative rates of peptide bond hydrolysis of the oxidized B chain of bovine insulin by two hemorrhagic proteinases, Ht-a and Ht-b, isolated from the venom of the Western Diamondback Rattlesnake, Crotalus atrox, were investigated. The results were compared with previous results on the digestion of the same substrate by the C. atrox hemorrhagic proteinases Ht-c, d, and e. Both toxins, Ht-a and Ht-b, were found to cleave the His5-Leu6, His10-Leu11, Ala14-Leu15, and Tyr16-Leu17 bonds of the oxidized insulin B chain. In addition, Ht-a cleaves the Asn3-Gln4 bond, whereas Ht-b cleaves the Gly23-Phe24 bond. The cleavage specificity of Ht-b on the insulin B chain is identical to that of the weakly hemorrhagic isoenzymes Ht-c and Ht-d. Hemorrhagic proteinase Ht-a cleaves the Ala14-Leu15 bond most rapidly, having a turnover number of 12.6 min-1 which was approximately twice as fast as the Tyr16-Leu17 bond turnover of cleavage. Hemorrhagic proteinase Ht-b, on the other hand, cleaves the Tyr16-Leu17 bond fastest, having a turnover number of 61.3 min-1. The Ala14-Leu15 bond is the peptide bond cleaved the second fastest by Ht-b, with a turnover number of 48.2 min-1. The five hemorrhagic toxins from Crotalus atrox venom were also examined for their capability to hydrolyse basement membrane preparations, and the identities of the digested proteins comprising the basement membrane were determined. From the results it is concluded that all five hemorrhagic toxins cleave laminin and the component of the basement membrane referred to as band a.(ABSTRACT TRUNCATED AT 250 WORDS)
Bjarnason
J B
JB
University of Iceland, Science Institute, Reykjavik.
Hamilton
D
D
Fox
J W
JW
D000595:Amino Acid Sequence; D000818:Animals; D001485:Basement Membrane / Q000378:metabolism; D003435:Crotalid Venoms / Q000633:toxicity*; D005337:Fibrin / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D006470:Hemorrhage / Q000139:chemically induced*; D006868:Hydrolysis; D007328:Insulin / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases
D003435:Crotalid Venoms; D005353:Fibronectins; D007328:Insulin; D007797:Laminin; D005337:Fibrin; D010447:Peptide Hydrolases