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PMID: 1904842 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Infection and immunity ·Vol. 59 ·No. 7 ·1991-07-00 ·Pages 2382-8

Geoffroy C, Raveneau J, Beretti JL, Lecroisey A, Vazquez-Boland JA, Alouf JE, Berche P

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.

MeSH Terms
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,isolation & purification,metabolism Zinc/metabolism
Chemicals
Hemolysin Proteins Metalloproteins Phosphatidylcholines Sphingomyelins Type C Phospholipases Zinc
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Geoffroy C
Unité des Antigènes Bactériens, Centre National de la Recherche Scientifique, Institut Pasteur, Paris, France.
Raveneau J
Beretti J L
Lecroisey A
Vazquez-Boland J A
Alouf J E
Berche P
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.59.7.2382-2388.1991
Published
1991-07-00
Pages
2382-8
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC258022
Subset
IM
External Links
PubMed source DOI: 10.1128/iai.59.7.2382-2388.1991 PMC Full Text View journal details
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