Characterization of Bothrops jararaca coagulation inhibitor (BjI) and presence of similar protein in plasma of other animals.
BjI, a protein isolated from Bothrops jararaca snake blood, inhibits the coagulant activity of thrombin. This protein presents two bands of 109 and 138 kDa by SDS-PAGE under reducing conditions. In order to verify the presence of BjI-like proteins in plasma of other animals (reptiles and non-reptiles), we raised a specific polyclonal antibody in mice to it, and we verified immunological cross-reaction by western blotting, considering as positive reactions the development of bands with either 109 or 138 kDa. Similar proteins were identified in Bothrops neuwiedi and Crotalus durissus terrificus snakes. In contrast, no BjI-like protein in other classes of animals was noticeable, nor in other snakes tested. Interestingly, a prolonged thrombin time was found only in snake plasmas that showed similar BjI proteins. BjI bound to two proteins of B. jararaca venom, identified by western blotting. The N-terminal of the B. jararaca venom proteins showed similarity with thrombin-like proteins isolated from other snake venoms. In conclusion, there are similar proteins to BjI in plasmas of B. neuwiedi and Crotalus durissus terrificus, and these proteins also prolong thrombin time. Moreover, these results evidence the presence of target enzymes in snake venom for plasma BjI.
Laboratório de Fisiopatologia, Instituto Butantan, Avenida Vital Brazil, 1500, CEP 05503-900 São Paulo SP, Brazil. amt.azevedo@uol.com.br
Anita M
AM
Tanaka-Azevedo
Ricardo J S
RJ
Torquato
Aparecida S
AS
Tanaka
Ida S
IS
Sano-Martins
D000595:Amino Acid Sequence; D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects*; D001798:Blood Proteins / Q000235:genetics* / Q000378:metabolism / Q000633:toxicity; D015153:Blotting, Western; D017837:Bothrops / Q000378:metabolism*; D001938:Brazil; D003429:Cross Reactions / Q000276:immunology; D003435:Crotalid Venoms / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D009943:Organophosphorus Compounds; D020539:Sequence Analysis, Protein; D013918:Thrombin Time
C043517:2-(4-isothiocyanatophenoxy)-1,3,2-dioxaphosphinene 2-oxide; D001798:Blood Proteins; C477871:Bothrops jararaca inhibitor; D003435:Crotalid Venoms; D007106:Immune Sera; D009943:Organophosphorus Compounds