Galleria mellonella native and analogue peptides Gm1 and ΔGm1. I) Biophysical characterization of the interaction mechanisms with bacterial model membranes

• Published in: Biochimica et biophysica acta 2014-10, Vol.1838 (10), p.2728-2738
• Main Authors: Correa, Wilmar, Manrique-Moreno, Marcela, Patiño, Edwin, Peláez-Jaramillo, Carlos, Kaconis, Yani, Gutsmann, Thomas, Garidel, Patrick, Heinbockel, Lena, Brandenburg, Klaus
• Format: Article
• Language: English
• Subjects: Animals; Anti-Infective Agents - chemistry; Antimicrobial Cationic Peptides - chemistry; Antimicrobial peptide; Attenuated total reflection; Bacteria - chemistry; Cell membrane; Cell Membrane - chemistry; Differential scanning calorimetry; Dimyristoylphosphatidylcholine - chemistry; Insect Proteins - chemistry; Lipopolysaccharide; Membranes, Artificial; Models, Chemical; Moths; Peptide–lipid interaction
• ISSN: 0005-2736; 0006-3002; 1879-2642
• DOI: 10.1016/j.bbamem.2014.07.006

Natural occurring antimicrobial peptides (AMPs) are important components of the innate immune system of animals and plants. They are considered to be promising alternatives to conventional antibiotics. Here we present a comparative study of two synthetic peptides: Gm1, corresponding to the natural overall uncharged peptide from Galleria mellonella (Gm) and ΔGm1, a modified overall positively charged Gm1 variant. We have studied the interaction of the peptides with lipid membranes composed of different kinds of lipopolysaccharides (LPS) and dimyristoylphosphatidylglycerol (DMPG), in some cases also dimyristoylphosphatidylethanolamine (DMPE) as representative lipid components of Gram-negative bacterial membranes, by applying Fourier-transform infrared spectroscopy (FTIR), Förster resonance energy transfer spectroscopy (FRET), differential scanning calorimetry (DSC) and isothermal titration calorimetry (ITC). Gm1 generates a destabilizing effect on the gel to liquid crystalline phase transition of the acyl chains of the lipids, as deduced from a decrease in the phase transition temperature and enthalpy, suggesting a fluidization, whereas ΔGm1 led to the opposite behavior. Further, FTIR analysis of the functional groups of the lipids participating in the interaction with the peptides indicated a shift in the band position and intensity of the asymmetric PO2− stretching vibration originating from the lipid phosphate groups, a consequence of the sterical changes in the head group region. Interestingly, FRET spectroscopy showed a similar intercalation of both peptides into the DMPG and LPS, but much less into the DMPE membrane systems. These results are discussed in the light of a possible use of the peptides as antimicrobial and anti-endotoxin drugs. [Display omitted] •Significant changes of the secondary structures of the peptides were observed.•The peptides influence the phase transition behavior of the lipids and changes in Tm.•Stronger changes in the phase transition enthalpy were detected.•The intercalation of peptides into phospholipids and lipopolysaccharides was similar.•The binding of the peptides to the LPS was enthalpically and entropically driven.