Effect of lactoferricin B, a pepsin-generated peptide of bovine lactoferrin, on Escherichia coli HB101 (pRI203) entry into HeLa cells.
Bovine lactoferrin and a pepsin-generated peptide of bovine lactoferrin, known as lactoferricin B, were tested for an ability to influence the cell-invasive properties of an Escherichia coli HB101 strain carrying the plasmid pRI203, which encodes the Yersinia pseudotuberculosis inv gene. At non-cytotoxic and non-bactericidal concentration (0.5 mg/ml) lactoferricin B lowered by about tenfold the cell invasion capability of E. coli HB101 (pRI203), whereas no effect was observed when bovine lactoferrin was added during the infection of HeLa cell monolayers. The step of the invasion process affected by lactoferrin B was the internalization since the adhesion of bacteria to HeLa cells was unaltered in the presence of the peptide. Latex beads coated with lactoferrin B bound to HeLa cell monolayers and induced the agglutination of bacterial cells, indicating that this highly cationic peptide interacts directly with both eukaryotic and bacterial surfaces. Moreover, we demonstrated that the anti-invasive effect induced by lactoferrin B was reversed when the medium was supplemented with Ca2+, Mg2+ and Fe2+ ions which diminished its affinity binding. Our findings suggest that lactoferrin B effectiveness towards E. coli HB101 (pRI203) invasion is correlated to its binding capability on the eukaryotic and bacterial cell surfaces.
Longhi
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C
Institute of Microbiology, Faculty of Medicine, University of Rome La Sapienza, Italy.
Conte
M P
MP
Bellamy
W
W
Seganti
L
L
Valenti
P
P
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001422:Bacterial Adhesion / Q000187:drug effects; D002413:Cations, Divalent / Q000494:pharmacology; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism / Q000472:pathogenicity*; D006367:HeLa Cells; D006801:Humans; D007781:Lactoferrin / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D008863:Microspheres; D010957:Plasmids; D015011:Yersinia pseudotuberculosis / Q000235:genetics
D000900:Anti-Bacterial Agents; D002413:Cations, Divalent; C082171:lactoferricin B; D007781:Lactoferrin