Purification and characterization of an extracellular 29-kilodalton phospholipase C from Listeria monocytogenes

Christiane Geoffroy, Jacques Raveneau, Jean-Luc Beretti, Anne Lecroisey, Jose A Vazquez-Boland, Joseph E Alouf, Patrick Berche

Research output: Contribution to journalArticlepeer-review

Abstract

We purified and characterized an extracellular phospholipase produced by Listeria monocytogenes. This enzyme was separated as a homogeneous protein of 29 kDa by chromatography on DEAE-52 cellulose and Bio-Gel P100 columns. It is a zinc-dependent phospholipase C (PLC) that is mainly active at pH 6 to 7 and expresses lecithinase activity and a weaker sphingomyelinase activity. The exoenzyme also hydrolyzed phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and sphingomyelin but not phosphatidylinositol. It was distinct from the 36-kDa phosphatidylinositol PLC produced by L. monocytogenes and from the L. ivanovii sphingomyelinase. The pure protein expressed a weak, calcium-independent hemolytic activity and was not toxic in mice. Western immunoblot analysis using a rabbit immune serum raised against the enzyme showed that all virulent strains of L. monocytogenes tested produced in the culture supernatant a 29-kDa PLC. In contrast, no proteins antigenically related to the 29-kDa PLC were detected in supernatants of L. ivanovii, L. seeligeri, L. innocua, or L. welshimeri. The role in virulence of the 29-kDa PLC specifically produced by L. monocytogenes remains to be established.
Original languageEnglish
Pages (from-to)2382-8
Number of pages7
JournalInfection and Immunity
Volume59
Issue number7
Publication statusPublished - Jul 1991

Keywords

  • Amino Acid Sequence
  • Blotting, Western
  • Extracellular Space/enzymology
  • Hemolysin Proteins
  • Hydrogen-Ion Concentration
  • Listeria monocytogenes/enzymology
  • Metalloproteins/metabolism
  • Molecular Sequence Data
  • Molecular Weight
  • Phosphatidylcholines/metabolism
  • Sphingomyelins/metabolism
  • Substrate Specificity
  • Type C Phospholipases/chemistry
  • Type C Phospholipases/isolation & purification
  • Type C Phospholipases/metabolism
  • Zinc/metabolism

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