Membrane topologies of the PGLa antimicrobial peptide and a transmembrane anchor sequence by Dynamic Nuclear Polarization/solid-state NMR spectroscopy.
Fiche publication
Date publication
février 2016
Journal
Scientific reports
Auteurs
Membres identifiés du Cancéropôle Est :
Pr BECHINGER Burkhard
Tous les auteurs :
Salnikov ES, Aisenbrey C, Aussenac F, Ouari O, Sarrouj H, Reiter C, Tordo P, Engelke F, Bechinger B
Lien Pubmed
Résumé
Dynamic Nuclear Polarization (DNP) has been introduced to overcome the sensitivity limitations of nuclear magnetic resonance (NMR) spectroscopy also of supported lipid bilayers. When investigated by solid-state NMR techniques the approach typically involves doping the samples with biradicals and their investigation at cryo-temperatures. Here we investigated the effects of temperature and membrane hydration on the topology of amphipathic and hydrophobic membrane polypeptides. Although the antimicrobial PGLa peptide in dimyristoyl phospholipids is particularly sensitive to topological alterations, the DNP conditions represent well its membrane alignment also found in bacterial lipids at ambient temperature. With a novel membrane-anchored biradical and purpose-built hardware a 17-fold enhancement in NMR signal intensity is obtained by DNP which is one of the best obtained for a truly static matrix-free system. Furthermore, a membrane anchor sequence encompassing 19 hydrophobic amino acid residues was investigated. Although at cryotemperatures the transmembrane domain adjusts it membrane tilt angle by about 10 degrees, the temperature dependence of two-dimensional separated field spectra show that freezing the motions can have beneficial effects for the structural analysis of this sequence.
Mots clés
Amino Acid Sequence, Anti-Infective Agents, chemistry, Antimicrobial Cationic Peptides, chemistry, Cold Temperature, Dimyristoylphosphatidylcholine, chemistry, Hydrophobic and Hydrophilic Interactions, Lipid Bilayers, chemistry, Magnetic Resonance Spectroscopy, instrumentation, Phosphatidylcholines, chemistry, Phosphatidylglycerols, chemistry
Référence
Sci Rep. 2016 Feb;6:20895