Membrane helix orientation from linear dichroism of infrared attenuated total reflection spectra.
Oriented multilamellar systems containing phospholipids and peptides have been formed on a germanium internal reflection element. Attenuated total reflection infrared spectra have been recorded and the linear dichroism of peptide amide I and amide II bands measured. Using peptides for which the orientation had been previously studied under similar experimental conditions by 15N solid-state nuclear magnetic resonance spectroscopy, important conclusions were drawn on the approach to be used to derive secondary structure orientation in a membrane from dichroic ratios. In particular, it is shown that the influence of the film thickness and refractive index on the orientation determination can be evaluated from the value of RATRiso, i.e., the dichroic ratio of a dipole oriented at the magic angle or with isotropic mobility. A series of peptides was used to test the validity of our suggestions on various helix orientations in the membrane. These include magainin 2 and hydrophobic (hPhi20) model peptides, the transmembrane segment of glycophorin (GLY), and LAH4, a designed peptide antibiotic that changes between a transmembrane and an in-plane orientation in a pH-dependent manner.
Bechinger
B
B
Max-Planck-Institut für Biochemie, Martinsried, Germany.
Ruysschaert
J M
JM
Goormaghtigh
E
E
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000737:chemistry; D023181:Antimicrobial Cationic Peptides; D055592:Biophysical Phenomena; D001703:Biophysics; D006021:Glycophorins / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D054806:Magainins; D008563:Membrane Lipids / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D017433:Protein Structure, Secondary; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D029867:Xenopus Proteins
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D006021:Glycophorins; D054806:Magainins; D008563:Membrane Lipids; D008565:Membrane Proteins; D010455:Peptides; D029867:Xenopus Proteins; C053301:magainin 2 peptide, Xenopus