Orientation and peptide-lipid interactions of alamethicin incorporated in phospholipid membranes: polarized infrared and spin-label EPR spectroscopy.
Alamethicin is a 20-residue peptaibiotic that induces voltage-dependent ion channels in lipid membranes. The mode by which alamethicin inserts into membranes was investigated using measurements of peptide-lipid interactions by spin-label electron paramagnetic resonance (EPR) and of peptide orientation by polarized infrared (IR) spectroscopy. In fluid membranes, spin-labeled stearic acid shows no evidence of a specific motionally restricted population of lipid chains, such as that found at the intramembranous surface of integral membrane proteins or oligomeric assemblies of transmembrane alpha-helices. In agreement with recent results from TOAC-substituted alamethicin analogues, native alamethicin is predominantly monomeric in fluid lipid membranes and presents an intramembrane surface that integrates well with the lipid chains but is insufficiently extensive to induce specific motional restriction. Channel formation takes place by transient association of transmembrane monomers. In aligned fluid membranes, alamethicin exhibits a large tilt in short chain-length lipids that decreases first rapidly with increasing chain-length and then more gradually for the lipids with longer chains. This macroscopically low order contrasts with the high local order, relative to the local membrane normal, that is found by EPR for alamethicins spin-labeled with TOAC. The macroscopic behavior is consistent with predictions for the chain-length dependence of elastic bending fluctuations of the membrane surface, which was invoked recently to explain the spontaneous insertion of beta-barrel proteins in short-chain lipid membranes.
Marsh
Derek
D
Max-Planck-Institut fur biophysikalische Chemie, Abt. Spektroskopie, 37070 Göttingen, Germany. dmarsh@gwdg.de
D000408:Alamethicin / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D008560:Membrane Fluidity / Q000502:physiology; D010455:Peptides / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D010713:Phosphatidylcholines / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D010743:Phospholipids / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D013055:Spectrophotometry, Infrared / Q000379:methods; D013113:Spin Labels; D019097:Spin Trapping; D014242:Trichoderma / Q000737:chemistry / Q000378:metabolism
D010455:Peptides; D010713:Phosphatidylcholines; D010743:Phospholipids; D013113:Spin Labels; D000408:Alamethicin