The antibiotic and DNA-transfecting peptide LAH4 selectively associates with, and disorders, anionic lipids in mixed membranes.
The histidine-rich amphipathic peptide LAH4 has antibiotic and DNA delivery capabilities. The peptide has a strong affinity for anionic lipids found in the outer membrane of bacterial membranes. A role for anionic lipids in release of cationic plasmid-containing complexes has been proposed previously, and disruption of membrane asymmetry and presentation of phosphatidylserine (PS) in the membrane outer leaflet is a general feature observed in diseased mammalian cells. Therefore, to understand the peptide-lipid interactions in more detail, solid-state NMR experiments on model membranes have been performed. 31P MAS NMR on mixed phosphatidylcholine (PC)/PS and PC/phosphatidylglycerol (PG) membranes has been used to demonstrate a strong interaction between LAH4 and anionic lipids. By using deuterated lipids and wide-line 2H NMR when probing lipid chain order, it is demonstrated that LAH4 preferentially interacts with PS over PC and effectively disorders the anionic PS lipid fatty acyl chains. In addition, we demonstrate that the efficiency of gene transfer in vitro to different cell lines is closely related to the degree of disruption of PS acyl chains for four isomers of LAH4. This work suggests a mechanism of selective destabilization by LAH4 of anionic lipids in the membranes of cells during transfection with implications for nucleic acid delivery in vivo.
Mason
A James
AJ
Faculté de chimie, University Louis Pasteur/CNRS UMR717-LC3, Institut le Bel, Strasbourg, France. jmason@chimie.u-strasbg.fr
Martinez
Amélie
A
Glaubitz
Clemens
C
Danos
Olivier
O
Kichler
Antoine
A
Bechinger
Burkhard
B
D000595:Amino Acid Sequence; D000838:Anions / Q000737:chemistry / Q000378:metabolism; D023181:Antimicrobial Cationic Peptides; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism; D004247:DNA; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008563:Membrane Lipids / Q000737:chemistry* / Q000378:metabolism*; D008567:Membranes, Artificial; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism; D013379:Substrate Specificity; D014162:Transfection
D000838:Anions; D023181:Antimicrobial Cationic Peptides; C122154:LAH(4) protein, synthetic; D008563:Membrane Lipids; D008567:Membranes, Artificial; D010455:Peptides; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D004247:DNA; D002784:Cholesterol; C028694:1-palmitoyl-2-oleoylphosphatidylcholine