Antibacterial and cell penetrating effects of LFcin17-30, LFampin265-284, and LF chimera on enteroaggregative Escherichia coli.
Lactoferrin (LF) is a protein with antimicrobial activity, which is conferred in part by 2 regions contained in its N-terminal lobe. These regions have been used to develop the following synthetic peptides: lactoferricin17-30, lactoferrampin265-284, and LF chimera (a fusion of lactoferricin17-30 and lactoferrampin265-284). We have reported that these LF peptides have antibacterial activity against several pathogenic bacteria; however, the exact mechanism of action has not been established. Here, we report the effects of LF peptides on the viability of enteroaggregative Escherichia coli (EAEC) and the ability of these peptides to penetrate into the bacteria cytoplasm. The viability of EAEC treated with LF peptides was determined via enumeration of colony-forming units, and the binding and internalization of the LF peptides was followed via immunogold labeling and electron microscopy. Treatment of EAEC with 20 and 40 μmol/L LF peptides reduced bacterial growth compared with untreated bacteria. Initially the peptides associated with the plasma membrane, but after 5 to 30 min of incubation, the peptides were found in the cytoplasm. Remarkably, bacteria treated with LF chimera developed cytosolic electron-dense structures that contained the antimicrobial peptide. Our results suggest that the antibacterial mechanism of LF peptides on EAEC involves their interaction with and penetration into the bacteria.
a CIASaP, Facultad de Medicina, Universidad Autónoma de Sinaloa, Cedros y Sauces, Fracc. Fresnos C.P. 80246, Culiacán, Sinaloa, Mexico.
Ruth
R
Reyes-Cortes
a CIASaP, Facultad de Medicina, Universidad Autónoma de Sinaloa, Cedros y Sauces, Fracc. Fresnos C.P. 80246, Culiacán, Sinaloa, Mexico.
Erika
E
Acosta-Smith
b Departamento de Bioquímica, Centro de Investigación y Estudios Avanzados del IPN, Avenida IPN #2508, Del. G.A. Madero, Ciudad de México, Mexico.
Ricardo
R
Mondragón-Flores
c Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam, University of Amsterdam and VU University, 1081 LA, Amsterdam, the Netherlands.
Kamran
K
Nazmi
c Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam, University of Amsterdam and VU University, 1081 LA, Amsterdam, the Netherlands.
Jan G M
JG
Bolscher
a CIASaP, Facultad de Medicina, Universidad Autónoma de Sinaloa, Cedros y Sauces, Fracc. Fresnos C.P. 80246, Culiacán, Sinaloa, Mexico.|d Departamento de Investigación, Hospital de la Mujer, Boulevard Miguel Tamayo Espinoza de los Monteros S/N, Col. Desarrollo Urbano Tres Ríos, Culiacán 80020, Sinaloa, Mexico.
Adrian
A
Canizalez-Roman
a CIASaP, Facultad de Medicina, Universidad Autónoma de Sinaloa, Cedros y Sauces, Fracc. Fresnos C.P. 80246, Culiacán, Sinaloa, Mexico.|e Departamento de Investigación, Hospital Pediátrico de Sinaloa, Boulevard Constitución S/N, col. Jorge Almada, Culiacan 80200, Sinaloa, Mexico.
Nidia
N
Leon-Sicairos
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D057846:Cell-Penetrating Peptides / Q000494:pharmacology*; D004926:Escherichia coli / Q000187:drug effects* / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000188:drug therapy* / Q000382:microbiology; D006801:Humans; D007781:Lactoferrin / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D057846:Cell-Penetrating Peptides; D007781:Lactoferrin
EAEC; LF chimera; LF peptides; antibacterial; antibactérien; chimère LF; lactoferrin; lactoferrine; peptides LF