Purification of a proteolytic enzyme from the venom os Bothrops brazili sanke and estudy of its activity on fibrinogen
A proteolitic enzyme was purified from Bothrops brazili peruvian snake venom using Sephadex G-100 followed by CM-Sephadex C-50, in both two cases with 0.05M ammonium acetate pH 7,0. The enzyme was purified 3,2 fold with 52,5% of yield and by gel filtration the enzyme showed 18 000 of molecular weight, while the PAGE-SIDS showed only one protein band of 22 000 in the presence of mercaptoethanol and 20 300 under nonreducing conditions which suggests that the enzyme has a single polypeptide chain with disulfide bond. The enzyme hydrolizes fibrinogen, fibrin, casein and albumin, but not hemoglobin and mioglobin. The enzyme is a Aa-fibrinogenase because preferentially hydrolizes the Aa chain of the fibrinogen molecule. This activity is inhibited by EDTA but not by PMSF, TLCK, iodoacetate and pepstatin, suggests that is a metalloproteinase, however the ions Ca++, Mg++ and Zn++ cannot reactivate the inhibition by EDTA. Finally the enzyme is stable up to 45 °C and in its effect on fibrin the enzyme is to attack a chain rapidly.
| Main Authors: | , , |
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| Format: | Digital revista |
| Language: | spa |
| Published: |
Universidad Nacional Mayor de San Marcos, Facultad de Ciencias Biológicas
2000
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| Online Access: | https://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/6728 |
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