LL-37observational2020

Membrane interactions of antimicrobial peptide-loaded microgels.

Journal of colloid and interface science

confidence

Key findings

LL-37 membrane interactions dominated by released peptide; microgel-loaded LL-37 showed sustained release, affecting peptide localization and bilayer structure.

View source on PubMed (PMID 31855795) ↗

Sample size
Not reported
Population
In vitro lipid membrane model (neutron reflectometry, FTIR-ATR, ellipsometry, CD experiments)
Dosing
LL-37 at 0.3 and 5.0 µM; microgel-loaded LL-37
Duration
Not reported
Route
In vitro
Blinding
not_reported
Controls
none
Drug class
antimicrobial peptide
Full abstract

In the present study, lipid membrane interactions of anionic poly(ethyl acrylate-co-methacrylic acid) (MAA) microgels as carriers for the cationic antimicrobial peptide LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES) were investigated. In doing so, neutron reflectometry (NR), Fourier-transform infrared spectroscopy with attenuated total reflection (FTIR-ATR), zeta potential, ellipsometry, and circular dichroism spectroscopy (CD) experiments were employed to investigate the relative importance of membrane interactions of peptide-loaded microgel particles and of released peptide. For the free peptide, NR results showed membrane binding occurring preferentially in the tail region in a concentration-dependent manner. At low peptide concentration (0.3 μM) only peptide insertion in the outer leaflet was seen, however, pronounced membrane defects and peptide present in both leaflets was observed at higher peptide concentration (5.0 μM). LL-37 loaded into MAA microgels qualitatively mirrored these effects regarding both peptide localization within the membrane and concentration-dependent defect formation. In addition, very limited membrane binding of microgel particles was observed, in agreement with FTIR-ATR and liposome leakage results. FTIR-ATR showed LL-37 to undergo α-helix formation on membrane insertion, also supported by CD results, the kinetics of which was substantially reduced for microgel-loaded LL-37 due to sustained peptide release. Together, these findings demonstrate that membrane interactions for microgel-loaded LL-37 are dominated by released peptide, but also that slow release of microgel-loaded LL-37 translates into kinetic effects on peptide-membrane interactions, relating to both peptide localization within the bilayer, and to bilayer structure.

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