Purification and characterization of an extracellular 29-kilodalton phospholipase C from Listeria monocytogenes.
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.
Geoffroy
C
C
Unité des Antigènes Bactériens, Centre National de la Recherche Scientifique, Institut Pasteur, Paris, France.
Raveneau
J
J
Beretti
J L
JL
Lecroisey
A
A
Vazquez-Boland
J A
JA
Alouf
J E
JE
Berche
P
P
D000595:Amino Acid Sequence; D015153:Blotting, Western; D005110:Extracellular Space / Q000201:enzymology; D006460:Hemolysin Proteins; D006863:Hydrogen-Ion Concentration; D008089:Listeria monocytogenes / Q000201:enzymology*; D008667:Metalloproteins / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010713:Phosphatidylcholines / Q000378:metabolism; D013109:Sphingomyelins / Q000378:metabolism; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D015032:Zinc / Q000378:metabolism
D006460:Hemolysin Proteins; D008667:Metalloproteins; D010713:Phosphatidylcholines; D013109:Sphingomyelins; D010738:Type C Phospholipases; D015032:Zinc