title: Conformation and membrane orientation of amphiphilic helical peptides by oriented circular dichroism.
abstract: Oriented circular dichroism (OCD) was used to characterize and compare in a quantitative manner the secondary structure and concentration dependent realignment of the antimicrobial peptides PGLa and MSI-103, and of the structurally related cell-penetrating peptide MAP in aligned phospholipid bilayers. All these peptides adopt an amphiphilic alpha-helical conformation, and from solid-state NMR analysis they are known to bind to membranes in two distinct orientations depending on their concentration. At low peptide/lipid (P/L) ratio the helices are aligned parallel to membrane surface (S-state), but with increasing concentration they realign to a tilted orientation (T-state), getting immersed into the membrane with an oblique angle supposedly as a result of dimer-formation. In macroscopically aligned liquid crystalline 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine bilayers the two limiting states are represented by distinct OCD spectra, and all spectra at intermediate peptide concentrations can be described by a linear combination of these two line shapes. The corresponding fraction of molecules occupying the T-state was determined by fitting the intermediate spectra with a superposition of the two extreme line shapes. By plotting this fraction versus 1/(P/L), the threshold P/L* ratio for realignment was extracted for each of the three related peptides. Despite their structural similarity distinctly different thresholds were obtained, namely for MSI-103 realignment starts already at a low P/L of approximately 1:236, for a MAP derivative (using a nonaggregating analog containing a D-amino acid) the transition begins at P/L approximately 1:156, whereas PGLa needs the highest concentration to flip into T-state at P/L approximately 1:85. Analysis of the original MAP sequence (containing only L-amino acids) gave OCD spectra compatible with beta-pleated conformation, suggesting that this peptide starts to aggregate with increasing concentration, unlike the other helical peptides. All these changes in peptide conformation and membrane alignment observed here by OCD seem to be functionally relevant, as they can be correlated with the membrane perturbing activities of the three antimicrobial and cell-penetrating sequences.
authors.affiliation: Karlsruhe Institute of Technology, Institute for Biological Interfaces (IBG-2), Karlsruhe, Germany.
authors.forename: Jochen
authors.initials: J
authors.lastname: Bürck
authors.forename: Siegmar
authors.initials: S
authors.lastname: Roth
authors.forename: Parvesh
authors.initials: P
authors.lastname: Wadhwani
authors.forename: Sergii
authors.initials: S
authors.lastname: Afonin
authors.forename: Nathalie
authors.initials: N
authors.lastname: Kanithasen
authors.forename: E
authors.initials: E
authors.lastname: Strandberg
authors.forename: Anne S
authors.initials: AS
authors.lastname: Ulrich
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D002942:Circular Dichroism; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D008051:Lipid Bilayers / Q000737:chemistry*; D044367:Phase Transition; D017433:Protein Structure, Secondary
chemical_list: C528508:(KIAGKIA)3 peptide; D023181:Antimicrobial Cationic Peptides; D008051:Lipid Bilayers; C045866:peptide-Gly-Leu-amide; D004134:Dimyristoylphosphatidylcholine