Characterization of a hemorrhagic factor, LHF-I, isolated from the bushmaster snake (Lachesis muta muta) venom.
Hemorrhagic factor I (LHF-I) was previously purified from the venom of the bushmaster snake (Lachesis muta muta). In terms of biochemical and immunological properties, LHF-I is a glycoprotein (mol. wt 100,000, pI 4.7) consisting of two subunits; it loses its activity following mercaptoethanol treatment. LHF-I contains 0.7 g-atom zinc and 1.2 g-atom calcium per mole protein. The hemorrhagic and the proteinase activities are inhibited by EDTA; subsequent addition of Ca2+ or Mg2+ does not reverse the EDTA-induced inhibition of the hemorrhagic activity. The metalloenzyme does not hyrolyze arginine esters and is devoid of phospholipase A2 activity. It hydrolyzes the A alpha- > B beta-chain of fibrinogen without clot formation and hydrolyzes selectively the alpha-chain of fibrin, leaving the B beta- and tau-chains unaffected. Antibodies to the hemorrhagic factor in bushmaster venom were produced by immunizing rabbits with the purified protein. The antibody was purified by protein-A affinity chromatography. This antibody was also used to screen other Crotalinae venom samples for immunologically similar epitopes by ELISA assay. The purified antibody reacted only with LHF-I and two samples of bushmaster venom from different geographical locations.
Sánchez
E F
EF
Centro de Pesquisa e Desenvolvimento, Fundaçao Ezequiel Dias, Belo Horizonte, M.G., Brazil.
Costa
M I
MI
Chavez-Olortegui
C
C
Assakura
M T
MT
Mandelbaum
F R
FR
Diniz
C R
CR
D000818:Animals; D000918:Antibody Specificity; D002118:Calcium / Q000378:metabolism; D003435:Crotalid Venoms / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism* / Q000633:toxicity; D004492:Edetic Acid / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005342:Fibrinolysis / Q000187:drug effects; D006470:Hemorrhage / Q000139:chemically induced; D005779:Immunodiffusion; D007525:Isoelectric Focusing; D007928:Lethal Dose 50; D008081:Liposomes / Q000378:metabolism; D008297:Male; D051379:Mice; D008970:Molecular Weight; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D012911:Snakes; D015032:Zinc / Q000378:metabolism
D003435:Crotalid Venoms; D008081:Liposomes; C032814:hemorrhagin components (Crotalid); D004492:Edetic Acid; D015032:Zinc; D002118:Calcium