Purification of an acidic phospholipase A2 from Bothrops lanceolatus (fer de lance) venom: molecular and enzymatic properties.
The acidic phospholipase A2 from Bothrops lanceolatus venom has been purified by gel filtration on Sephadex G-50 and ion exchange chromatography on CM-cellulose. Analysis by FPLC on Mono-Q column of the purified phospholipase A2 indicated that it is a mixture of several isoenzymes. The two major isoforms consist of a single polypeptide chain with mol. wts of 14,500 and 15,000, which slightly differ in their isoelectric point (4.9 and 5.3) and amino acid composition. However, enzymatic and pharmacological properties of the various isoenzymes are identical. The phospholipase from B. lanceolatus venom is characterized by a progressive increase in the rate of hydrolysis when enzymatic activity is determined with crude egg yolk as substrate in the absence of detergent. This phenomenon, which is not observed with mixed micelles of lecithin-detergent, is not due to the presence of a phospholipase A2 inhibitor in the venom, as previously suggested by several investigators in the case of other Bothrops and Cobra venoms. It is rather a catalytic characteristics of B. lanceolatus venom phospholipase, the enzymatic activity of which depends on the physical state of phospholipids. Bothrops lanceolatus acidic phospholipase A2 is non-toxic.
de Araújo
A L
AL
Unité des Venins, Institut Pasteur, Paris, France.
Radvanyi
F
F
Bon
C
C
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D017837:Bothrops; D002645:Chickens; D002852:Chromatography, Ion Exchange; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000633:toxicity; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000302:isolation & purification; D008297:Male; D051379:Mice; D008970:Molecular Weight; D009026:Mortality; D010741:Phospholipases A / Q000737:chemistry / Q000302:isolation & purification*; D054467:Phospholipases A2
D000596:Amino Acids; D003435:Crotalid Venoms; D007527:Isoenzymes; D010741:Phospholipases A; D054467:Phospholipases A2