期刊
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
卷 1848, 期 6, 页码 1367-1375出版社
ELSEVIER
DOI: 10.1016/j.bbamem.2015.03.007
关键词
Antimicrobial peptide (AMP); Cell-penetrating peptide (CPP); Phospholipid vesicle; Membrane lipid; Drug resistance
To understand the role of cell membrane phospholipids during resistance development to cationic antimicrobial peptides (CAMPs) in Enterococcus faecalis, gradual dose-dependent single exposure pediocin-resistant (Ped(r)) mutants of E. faecalis (Efv2.1, Efv3.1, Efv3.2, Efv4.1, Efv4.2, Efv5.1, Efv5.2 and Efv5.3), conferring simultaneous resistance to other CAMPs, selected in previous study were characterized for cell membrane phospholipid head-groups and fatty acid composition. The involvement of phospholipids in resistance acquisition was confirmed by in vitro colorimetric assay using PDA (polydiacetylene)-biomimetic membranes. Estimation of ratio of amino-containing phospholipids to amino-lacking phospholipids suggests that phospholipids in cell membrane of Ped(r) mutants loose anionic character. At moderate level of resistance, the cell-membrane becomes neutralized while at further higher level of resistance, the cell-surface acquired positive charge. Increased expression of mprF gene (responsible for lysinylation of phospholipids) was also observed on acquiring resistance to pediocin in Ped(r) E. faecalis. Decreased level of branched chain fatty acids in Ped(r) mutants might have contributed in enhancing rigidification of cell membrane and contributing towards resistance. The interaction of pediocin with PDA-biomimetic membranes prepared from wild-type and Ped(r) mutants was monitored by measuring percent colorimetric response (%CR). Increased %CR of pediocin against PDA-biomimetic membranes prepared from Ped(r) mutants confirmed that cell membrane phospholipids are involved in the interactions of pore formation by CAMPs. There was a direct linear relationship between percent calorimetric response and IC50 of CAMPs for wild-type and Ped(r) mutants. This relationship further reveals that in vitro colorimetric assay can be used effectively for quantification of resistance to CAMPs. (C) 2015 Elsevier B.V. All rights reserved.
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