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Abstract
The objective of the present study was an investigation of the crude Bothrops moojeni venom, aiming at the identification of new compounds with platelet-activating or -inhibiting activity. The venom was separated by gel filtration chromatography into 18 fractions, which were tested by means of whole blood aggregometry for their activities affecting the aggregation of blood platelets. In order to eliminate interferences caused by prothrombin activators or thrombin like-enzymes, which are frequently present in snake venoms, a test method for screening protein mixtures was developed. To avoid clotting of the blood samples, the thrombin inhibitor hirudin and the synthetic inhibitor of fibrin polymerization Pefabloc® FG were applied. In the present study, a platelet aggregation activator with an activity resembling thrombocytin from B. atrox was identified in one of the examined venom fractions. In addition, a platelet antagonist–most likely a disintegrin–with broad inhibitory activity against aggregation triggered by collagen, adenosine diphosphate and thrombin receptor activating peptide, was identified.
| Abbreviations | ||
| ACN | = | acetonitrile |
| ADP | = | adenosine diphosphate |
| AU | = | aggregation unit |
| COL | = | collagen |
| FPI | = | fibrin polymerization inhibitor |
| GF | = | gel filtration |
| HIC | = | hydrophobic interaction chromatography |
| LAAO | = | L-amino acid oxidase |
| MALDI-TOF | = | matrix assisted laser desorption ionisation–time of flight |
| MS | = | mass spectrometry |
| MSP 1 and 2 | = | basic serine proteases from the venom of Bothrops moojeni |
| MW | = | molecular weight |
| PAR | = | protease-activated receptor |
| PA-BJ | = | platelet-aggregating enzyme from the venom of B. jararaca |
| PLA2 | = | phospholipase A2 |
| PRP | = | platelet-rich plasma |
| PTA | = | prothrombin activator |
| RT | = | room temperature |
| SPE | = | solid phase extraction |
| TFA | = | trifluoroacetic acid |
| TLE | = | thrombin-like enzyme |
| TRAP-6 | = | thrombin receptor activating peptide |
| vWF | = | von Willebrand factor. |
| Abbreviations | ||
| ACN | = | acetonitrile |
| ADP | = | adenosine diphosphate |
| AU | = | aggregation unit |
| COL | = | collagen |
| FPI | = | fibrin polymerization inhibitor |
| GF | = | gel filtration |
| HIC | = | hydrophobic interaction chromatography |
| LAAO | = | L-amino acid oxidase |
| MALDI-TOF | = | matrix assisted laser desorption ionisation–time of flight |
| MS | = | mass spectrometry |
| MSP 1 and 2 | = | basic serine proteases from the venom of Bothrops moojeni |
| MW | = | molecular weight |
| PAR | = | protease-activated receptor |
| PA-BJ | = | platelet-aggregating enzyme from the venom of B. jararaca |
| PLA2 | = | phospholipase A2 |
| PRP | = | platelet-rich plasma |
| PTA | = | prothrombin activator |
| RT | = | room temperature |
| SPE | = | solid phase extraction |
| TFA | = | trifluoroacetic acid |
| TLE | = | thrombin-like enzyme |
| TRAP-6 | = | thrombin receptor activating peptide |
| vWF | = | von Willebrand factor. |