Isolation by preparative isoelectric focusing of a direct acting fibrinolytic enzyme from the venom of Agkistrodon contortrix contortrix (southern copperhead).
Fibrolase, a blood clot-lysing enzyme, was isolated from the venom of the snake Agkistrodon contortrix contortrix using preparative scale isoelectric focusing in the recycling isoelectric focusing (RIEF) apparatus. Two sequential purifications, beginning with 1.0 g of whole, dried venom, were employed. A pH 6-8 range gradient effected the first separation. While 100% of the enzyme was recovered in three fractions, 43% (one fraction) had 70% purity. The second run was a refractionation of three, pooled fractions from the first run, in a 0.7 pH range gradient. Of the fibrolase in the venom, 63% was recovered in four fractions. One of these represented 29% of venom fibrolase, with 97% purity. Gel filtration chromatography removed most of the remaining, higher molecular weight contaminants of the RIEF-purified enzyme.
D000818:Animals; D002364:Caseins / Q000032:analysis; D002850:Chromatography, Gel; D003435:Crotalid Venoms / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D005342:Fibrinolysis; D007525:Isoelectric Focusing; D011492:Protein Hydrolysates / Q000032:analysis; D012834:Silver; D012967:Sodium Dodecyl Sulfate; D013194:Staining and Labeling
C031442:Agkistrodon venoms; D002364:Caseins; D003435:Crotalid Venoms; D011492:Protein Hydrolysates; C026375:trypticase-soy broth; D012967:Sodium Dodecyl Sulfate; D012834:Silver