Staphylococcal delta toxin stimulates endogenous phospholipase A2 activity and prostaglandin synthesis in fibroblasts

Jon P. Durkin, W. T. Shier

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

Delta toxin, one of at least four toxins produced by pathogenic strains of the skin bacterium Staphylococcus aureus, is an amphipathic polypeptide possessing hemolytic and cytolytic activity. Delta toxin stimulates high levels of phospholipase A2 activity in 3T3 mouse fibroblasts with concomitant synthesis and release of prostaglandins. Alpha toxin, another hemolytic toxin produced by strains of S. aureus, did not stimulate phospholipase A2 or prostaglandin release in these cells. Analysis of the release of lactate dehydrogenase and β-galactosidase (cytoplasmic and lysosomal marker enzymes, respectively) from delta-toxin-treated cells indicated that cytolytic concentrations of the toxin damage the cell-surface membrane more extensively than lysosomal membranes. During a 30 min exposure, delta toxin stimulated 3T3 cells to hydrolyze up to 32% of the lipids biosynthetically labeled by incorporation of [3H]arachidonic acid. A relatively high percentage of the free arachidonic acid formed in delta-toxin-treated 3T3 cells was converted to prostaglandins (up to 41.3% and 8.3% converted to chromatographically identifiable prostaglandins E2 and F, respectively, in 30 min), with optimal conversion occurring at sublytic toxin concentrations. The degree of activation of phospholipase A2 in 3T3 cells by a range of concentrations of delta toxin correlates with cytotoxicity assessed by failure to exclude trypan blue dye. Analysis of the calcium dependency of the toxin-activated phospholipase A2 was consistent with a cell-surface, Ca2+-dependent enzyme. The phospholipase A2 exhibits a degree of specificity for substrate lipids containing polyunsaturated fatty acid residues which can serve as precursors for prostaglandin formation. Enzymatic activity was not inhibited by diisopropylfluorophosphate (5 mM), N-ethylmaleimide (5 mM) or p-bromophenacylbromide (0.1 mM). Delta toxin did not activate detectable phospholipase A2 in subcellular preparations containing plasma membrane.

Original languageEnglish (US)
Pages (from-to)467-479
Number of pages13
JournalBiochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism
Volume663
Issue number2
DOIs
StatePublished - Feb 23 1981

Bibliographical note

Funding Information:
We gratefully acknowledge gifts of samples of staphylococcal toxins and S. aureus Wood 46M from Dr. A.W. Bernheimer and the technical assistanceo f Mr. J.T. Trotter. This research was supported in part by a fellowship from the National Research Council of Canada (to J.P.D.); a grant from the Cystic Fibrosis Foundation; Grant Number CA20636, awarded by the National Cancer Institute, D.H.E.W., and Grant Number PCM80-11784, awarded by the National Science Foundation.

Keywords

  • (Staphylococcus
  • Cytolysis
  • Delta toxin
  • Mouse fibroblast)
  • Phospholipase A
  • Prostaglandin synthesis

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