Purification and characterization of a lethal protein with phospholipase A1 activity from the hornet (Vespa basalis) venom.
The hornet, Vespa basalis, is one of the most dangerous species of wasps found in Taiwan. The insect is aggressive and its venom is highly toxic. By gel filtration on a Fractogel TSK HW 50 column followed by cation-exchange chromatography on CM-Trisacryl M, a lethal protein was purified from the venom. It has a molecular mass of about 32 kDa and an i.v. LD50 value of 0.32 micrograms/g mouse. The toxin is capable of catalyzing the hydrolysis of emulsified phospholipids but not sphingomyelin. Analysis of the 1H-NMR spectra of the substrates and its hydrolytic products revealed that the toxin liberates fatty acid from the 1-position of sn-3-phosphoacylglycerols. This result indicates that the toxin possesses phospholipase A1 activity. The toxin exhibits an extremely potent hemolytic activity in washed red cells and diluted whole blood (HC50 = 0.09 micrograms/ml in mouse). The potency of direct hemolysis is about 100-times that of a basic phospholipase A2 from Naja nigricollis venom and about 1000-times that of a cardiotoxin from Naja naja atra venom. A positive correlation between the hemolytic activity and lethality of the toxin was found in three species of animals (mouse, rat and guinea pig). In the in vivo study, the toxin caused a marked increase in the plasma K+ concentration and a hyperkalemic change in the ECG of the treated rat. Hyperkalemia resulting from the hemolytic action of the toxin appears to be the main cause of death in the animal.
Ho
C L
CL
Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan, China.
Ko
J L
JL
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001514:Bee Venoms / Q000032:analysis* / Q000633:toxicity; D001794:Blood Pressure / Q000187:drug effects; D002118:Calcium / Q000378:metabolism; D004546:Elapid Venoms / Q000633:toxicity; D004562:Electrocardiography; D006168:Guinea Pigs; D006461:Hemolysis; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008970:Molecular Weight; D010740:Phospholipases / Q000302:isolation & purification*; D010741:Phospholipases A / Q000302:isolation & purification* / Q000378:metabolism; D054466:Phospholipases A1; D054467:Phospholipases A2; D011188:Potassium / Q000097:blood; D051381:Rats; D013379:Substrate Specificity; D014862:Wasp Venoms / Q000032:analysis* / Q000633:toxicity
D000596:Amino Acids; D001514:Bee Venoms; D004546:Elapid Venoms; D014862:Wasp Venoms; C050935:hornetin; D010740:Phospholipases; D010741:Phospholipases A; D054466:Phospholipases A1; D054467:Phospholipases A2; D011188:Potassium; D002118:Calcium