Purification and partial characterization of a haemorrhagin (VRH-1) from Vipera russelli russelli venom.
A haemorrhagic toxin (VRH-1) has been purified to homogeneity from Vipera russelli russelli venom by subjecting it to chromatography twice successively on CM-Sephadex C-50. It is a protein of mol. wt 22,000 and contains one mole of Mg2+. Intradermal administration of this haemorrhagin in mice resulted in severe lung haemorrhage but produced little haemorrhage in skin. This apparent organ preference led us to develop a new haemorrhage assay method utilizing dye diffusion from lung in vitro. Proteolytic activity of VRH-1 was demonstrated using dimethylcasein as substrate following quantitation by reaction with trinitrobenzoyl sulfonic acid. Both haemorrhagic and proteolytic activities of VRH-1 were inhibited by serine protease inhibitors like phenylmethyl sulfonyl fluoride and chymostatin, but metal chelators had no effect. Lung haemorrhage is unlikely to be a direct reflection of a high local concentration of VRH-1. The administration of supernatant generated by incubation of chopped liver from untreated mouse and VRH-1 (in subhaemorrhagic dose) results in severe lung haemorrhage. This raises the possibility that VRH-1 leads to the formation of intermediate(s) which causes the haemorrhage.
Chakrabarty
D
D
Indian Institute of Chemical Biology, Calcutta.
Bhattacharyya
D
D
Sarkar
H S
HS
Lahiri
S C
SC
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006470:Hemorrhage / Q000097:blood / Q000139:chemically induced*; D066298:In Vitro Techniques; D008297:Male; D008670:Metals / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D011517:Prothrombin Time; D051381:Rats; D017840:Daboia / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D014018:Tissue Distribution; D014757:Viper Venoms / Q000737:chemistry* / Q000493:pharmacokinetics
D000596:Amino Acids; D002241:Carbohydrates; D008670:Metals; D011506:Proteins; D015842:Serine Proteinase Inhibitors; D014757:Viper Venoms; C086622:haemorrhagic toxin VRH-1