Purification and characterization of a phospholipase A2 isoenzyme isolated from Lachesis muta snake venom

Biochem Pharmacol. 2002 May 1;63(9):1589-97. doi: 10.1016/s0006-2952(02)00873-0.

Abstract

A new phospholipase A2 (PLA2) isoenzyme was isolated from Lachesis muta crude venom, and was named LM-PLA2-II. This enzyme was purified by gel filtration on a Sephacryl S-200 HR column followed by reverse-phase chromatography on a C2/C18 column. LM-PLA2-II consists of a single polypeptide chain with an apparent molecular mass of 18 kDa and an isoelectric point at pH 5.4. The amino terminal sequence of the enzyme revealed a high degree of homology with other PLA2s from several sources. LM-PLA2-II has a high indirect hemolytic activity and a potent inhibitory effect on platelet aggregation induced by ADP and collagen. It also produces a significant paw edema reaction in rats. The edematous response in rats was abolished by pretreatment with either indomethacin or dexamethasone, suggesting the involvement of cyclo-oxygenase. Pretreatment of LM-PLA2-II with p-bromophenacyl bromide abolished all of these actions, clearly indicating that the biological activities, including the edematogenic effect, are dependent entirely on its enzymatic activity.

MeSH terms

  • Adenosine Diphosphate / physiology
  • Amino Acid Sequence
  • Animals
  • Blood Platelets / drug effects
  • Blood Platelets / physiology
  • Collagen / physiology
  • Edema / chemically induced
  • Hemolysis / drug effects
  • Hemorrhage / chemically induced
  • Isoenzymes / isolation & purification*
  • Isoenzymes / metabolism
  • Isoenzymes / pharmacology
  • Molecular Sequence Data
  • Phospholipases A / isolation & purification*
  • Phospholipases A / metabolism
  • Phospholipases A / pharmacology
  • Phospholipases A2
  • Platelet Aggregation Inhibitors / isolation & purification
  • Platelet Aggregation Inhibitors / pharmacology
  • Sequence Homology, Amino Acid
  • Viper Venoms / enzymology*
  • Viperidae / metabolism*

Substances

  • Isoenzymes
  • Platelet Aggregation Inhibitors
  • Viper Venoms
  • Adenosine Diphosphate
  • Collagen
  • Phospholipases A
  • Phospholipases A2