Partial purification and some physicochemical properties of phospholipases A2 from the venom of the bushmaster snake (Lachesis muta).
Screening of the biochemical-pharmacological properties of the crude venom from the snake Lachesis muta indicated the presence of phospholipase A2 (PLA2; 5260 U/mg protein), procoagulant (2630 U/mg protein), platelet aggregating (43 U/mg protein) and caseinolytic activities (6670 U/mg protein). These activities were separated by filtration of the crude venom on Sephacryl S-200. The material containing PLA2 activity was further fractioned by DEAE-cellulose ion exchange chromatography into four active fractions (F-I to F-IV, containing 1.7, 1.2, 0.3, and 0.05% of the crude venom protein, respectively) by stepwise elution with buffers of increasing ionic strength. All fractions presented a molecular weight of approximately 15,000 and isoelectric points in the range pH 4.6-6.0. In addition to their indirect hemolytic activity, the partially purified fractions inhibited platelet aggregation induced either by collagen or thrombin. p-Bromophenacyl bromide-treated fractions lost both phospholipase A2 activity and their inhibitory effect on collagen-induced platelet aggregation.
Fuly
A L
AL
Departamento de Bioquímica, Universidade Federal do Rio de Janeiro, Brasil.
Francischetti
I M
IM
Zingali
R B
RB
Carlini
C R
CR
D000818:Animals; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D010741:Phospholipases A / Q000302:isolation & purification* / Q000494:pharmacology; D054467:Phospholipases A2; D010974:Platelet Aggregation; D014757:Viper Venoms / Q000737:chemistry* / Q000201:enzymology / Q000378:metabolism; D017819:Viperidae
D014757:Viper Venoms; D010741:Phospholipases A; D054467:Phospholipases A2