A Novel Acidic Myotoxic Phospholipase A2 of Colombian B. asper Venom. Isolation and Functional Characterization
نویسندگان
چکیده
Myotoxic phospholipases A2 are responsible for many clinical signs in envenomation by Bothrops snakes. A new myotoxic acidic PLA2 Asp 49 was isolated from Colombian Bothrops asper venom. It was isolated by high efficiency liquid chromatography and named BaCol PLA2. It had a molecular weight of 14180.69 Da and an isoelectric point of 4.4. The complete sequence obtained by cDNA cloning, with access number in the gene bank MF319968; this sequence evidenced a mature product of 124 amino acids with Asp in 49 position. In vivo assays in mice demonstrated profuse oedema and myotoxicity evidenced by increase of creatina kinasa in plasma and severe and diffuse damage to the muscular fibers, further vacuolization and hyalinization necrosis of the sarcoplasm showed by histopathology with hematoxylin and eosin staining of gastrocnemius muscle. In vitro studies showed cell membrane damage without phosphatidylserine exposure, an early apoptosis hallmark. Further BaCol PLA2 evidenced high indirect hemolytic activity and moderate anticoagulant action. The toxin showed homology with others acidic PLA2 isolated from Bothrops venoms, including one isolated from B. asper of Costa Rica. Unlike this, BaCol PLA2 was myotoxic.
منابع مشابه
Isolation and Functional Characterization of an Acidic Myotoxic Phospholipase A2 from Colombian Bothrops asper Venom
Myotoxic phospholipases A₂ (PLA₂) are responsible for many clinical manifestations in envenomation by Bothrops snakes. A new myotoxic acidic Asp49 PLA₂ (BaCol PLA₂) was isolated from Colombian Bothrops asper venom using reverse-phase high performance liquid chromatography (RP-HPLC). BaCol PLA₂ had a molecular mass of 14,180.69 Da (by mass spectrometry) and an isoelectric point of 4.4. The compl...
متن کاملIsolation and characterization of a myotoxic phospholipase A2 from the venom of the arboreal snake Bothriechis (Bothrops) schlegelii from Costa Rica.
A new myotoxic phospholipase A2 was isolated from the venom of the arboreal snake Bothriechis schlegelii (formerly Bothrops schlegelii) from Costa Rica, by ion-exchange chromatography on CM-Sephadex. B. schlegelii myotoxin I is a basic protein (pI > 9.3) with a subunit molecular weight of 15 kDa, which migrates as a dimer in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under nonred...
متن کاملSynergism between Basic Asp49 and Lys49 Phospholipase A2 Myotoxins of Viperid Snake Venom In Vitro and In Vivo
Two subtypes of phospholipases A2 (PLA2s) with the ability to induce myonecrosis, 'Asp49' and 'Lys49' myotoxins, often coexist in viperid snake venoms. Since the latter lack catalytic activity, two different mechanisms are involved in their myotoxicity. A synergism between Asp49 and Lys49 myotoxins from Bothrops asper was previously observed in vitro, enhancing Ca2+ entry and cell death when ac...
متن کاملEffects of Bothrops asper Snake Venom on Lymphatic Vessels: Insights into a Hidden Aspect of Envenomation
BACKGROUND Envenomations by the snake Bothrops asper represent a serious medical problem in Central America and parts of South America. These envenomations concur with drastic local tissue pathology, including a prominent edema. Since lymph flow plays a role in the maintenance of tissue fluid balance, the effect of B. asper venom on collecting lymphatic vessels was studied. METHODOLOGY/PRINCI...
متن کاملBiochemical characterization and pharmacological properties of a phospholipase A2 myotoxin inhibitor from the plasma of the snake Bothrops asper.
A protein that neutralizes the biological activities of basic phospholipase A2 (PLA2) myotoxin isoforms from the venom of the snake Bothrops asper was isolated from its blood by affinity chromatography with Sepharose-immobilized myotoxins. Biochemical characterization of this B. asper myotoxin inhibitor protein (BaMIP) indicated a subunit molecular mass of 23-25 kDa, an isoelectric point of 4, ...
متن کامل