Abstract
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 NHF a and NHF b. 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 NHF b is 23 times stronger than that of NHF a. Both hydrolyse casein, although NHF a is about 20 times more active than NHF b. They are metalloproteins inhibited by EDTA and 1,10-phenanthroline. NHF a and NHF b 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 NHF a from NHF b showing, nevertheless, that they have common determinants apart from specific determinants for each one.
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