Characterization of two hemorrhagic factors isolated from the venom of Bothrops neuwiedi (jararaca pintada)

• Published in: Toxicon (Oxford) Vol. 22; no. 2; p. 193
• Main Authors: Mandelbaum, F R, Assakura, M T, Reichl, A P
• Format: Journal Article
• Language: English
• Published: England 1984
• Subjects: Animals; Blood Coagulation - drug effects; Carboxylic Ester Hydrolases - analysis; Caseins; Crotalid Venoms - isolation & purification; Crotalid Venoms - toxicity; Electrophoresis, Polyacrylamide Gel - methods; Hemorrhage - chemically induced; Immunodiffusion; In Vitro Techniques; Isoelectric Focusing; Molecular Weight; Peptide Hydrolases - analysis; Proteins - analysis; Sheep
• ISSN: 0041-0101
• DOI: 10.1016/0041-0101(84)90020-5

Two hemorrhagic factors were isolated from the venom of Bothrops neuwiedi (jararaca pintada) by ammonium sulfate precipitation followed by chromatography on DEAE-Sephadex A-50 and DEAE-cellulose DE-32, gel filtration on Sephadex G-100 and polyacrylamide-gel electrophoresis. These factors were named neuwiedi hemorrhagic factors NHFa and NHFb. They are acidic proteins of pI 4.2-4.3 and consist of single polypeptide chains of molecular weights 46,000 and 58,000, respectively, as determined by sodium dodecyl sulfate polyacrylamide-gel electrophoresis. The hemorrhagic activity of NHFb is 23 times stronger than that of NHFa. Both hydrolyse casein, although NHFa is about 20 times more active than NHFb. They are metalloproteins inhibited by EDTA and 1,10-phenanthroline. NHFa and NHFb are serologically closely related antigens. These two factors are recognized as identical antigens by horse serum against crude Bothrops neuwiedi venom. However, the rabbit specific antiserum was able to differentiate NHFa from NHFb showing, nevertheless, that they have common determinants apart from specific determinants for each one.