Stimulation of cell surface phospholipase A2 and prostaglandin synthesis in 3T3 mouse fibroblasts by phallolysin, a toxin from Amanita phalloides

W. Thomas Shier, Joseph T. Trotter

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Phallolysin, a cytotoxic glycoprotein from the poisonous mushroom Amanita phalloides, stimulates high levels of cellular phospholipase A2 in 3T3 Swiss mouse fibroblasts biosynthetically labeled by incorporation of [5,6,8,9, 11,12,14,15-3H]arachidonic acid. Up to 39.9% of labeled cell lipids were hydrolyzed in 30 min with concomitant synthesis of prostaglandins (up to 5.3 and 1.7% of incorporated label released in 30 min as chromatographically identifiable prostaglandins E2 and F, respectively). A similar extent of hydrolysis of labeled phosphatidylcholine was observed in cells biosynthetically labeled with [1,2-14C]choline, with production of an equivalent amount of lysophosphatidylcholine. No detectable hydrolysis of sphingomyelin was observed, indicating (1) that phospholipases B, C and D and sphingomyelinases were not significantly activated, and (2) that the phospholipase A2 activated did not exhibit a high degree of specificity for arachidonic acid moieties in its substrates. An analysis of the Ca2+ requirement for activation suggests that phallolysin activates primarily cell surface, Ca2+-requiring phospholipase A2. Several lines of evidence suggest that stimulation of phospholipase A2 by phallolysin involves binding to receptor sites which are also sites for wheat germ agglutinin. The extent of cell death, as assessed by failure to exclude trypan blue dye, correlated with the percent of maximal activation of phospholipase A2 by a range of concentrations of phallolysin, suggesting that the toxin kills cells by inducing hydrolysis of membrane phospholipids.

Original languageEnglish (US)
Pages (from-to)235-246
Number of pages12
JournalBiochimica et Biophysica Acta - Lipids and Lipid Metabolism
Volume619
Issue number2
DOIs
StatePublished - 1980

Keywords

  • Cell surface
  • Fibroblast
  • Phallolysin
  • Phospholipase A
  • Prostaglandin
  • Toxin

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