Disruption of Saccharomyces cerevisiae by Plantaricin 149 and investigation of its mechanism of action with biomembrane model systems.
The action of a synthetic antimicrobial peptide analog of Plantaricin 149 (Pln149a) against Saccharomyces cerevisiae and its interaction with biomembrane model systems were investigated. Pln149a was shown to inhibit S. cerevisiae growth by more than 80% in YPD medium, causing morphological changes in the yeast wall and remaining active and resistant to the yeast proteases even after 24 h of incubation. Different membrane model systems and carbohydrates were employed to better describe the Pln149a interaction with cellular components using circular dichroism and fluorescence spectroscopies, adsorption kinetics and surface elasticity in Langmuir monolayers. These assays showed that Pln149a does not interact with either mono/polysaccharides or zwitterionic LUVs, but is strongly adsorbed to and incorporated into negatively charged surfaces, causing a conformational change in its secondary structure from random-coil to helix upon adsorption. From the concurrent analysis of Pln149a adsorption kinetics and dilatational surface elasticity data, we determined that 2.5 muM is the critical concentration at which Pln149a will disrupt a negative DPPG monolayer. Furthermore, Pln149a exhibited a carpet-like mechanism of action, in which the peptide initially binds to the membrane, covering its surface and acquiring a helical structure that remains associated to the negatively charged phospholipids. After this electrostatic interaction, another peptide region causes a strain in the membrane, promoting its disruption.
Lopes
José Luiz S
JL
Biofísica Molecular Sergio Mascarenhas, Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, SP, Brazil.
Nobre
Thatyane M
TM
Siano
Alvaro
A
Humpola
Verónica
V
Bossolan
Nelma R S
NR
Zaniquelli
Maria E D
ME
Tonarelli
Georgina
G
Beltramini
Leila M
LM
D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology; D001430:Bacteriocins / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D008051:Lipid Bilayers / Q000378:metabolism*; D008081:Liposomes / Q000378:metabolism; D008954:Models, Biological; D010743:Phospholipids / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D013500:Surface Tension
D023181:Antimicrobial Cationic Peptides; D001430:Bacteriocins; D008051:Lipid Bilayers; D008081:Liposomes; D010743:Phospholipids; C519206:plantaricin 149