title: Mapping the orientation of helices in micelle-bound peptides by paramagnetic relaxation waves.
abstract: Many antimicrobial peptides form alpha-helices when bound to a membrane. In addition, around 80% of residues in membrane-bound proteins are found in alpha-helical regions. The orientation and location of such helical peptides and proteins in the membrane are key factors determining their function and activity. Here we present a new solution state NMR method for obtaining the orientation of helical peptides in a membrane-mimetic environment (micelle-bound) without any chemical perturbation of the peptide-micelle system. By monitoring proton longitudinal relaxation rates upon addition of the freely water-soluble and inert paramagnetic probe Gd(DTPA-BMA) to an alpha-helical peptide, a wavelike pattern with a periodicity of 3.6 residues per turn is observed. The tilt and azimuth (rotation) angle of the helix determine the shape of this paramagnetic relaxation wave and can be obtained by least-square fitting of measured relaxation enhancements. Results are presented for the 15-residue antimicrobial peptide CM15 which forms an amphipathic helix almost parallel to the surface of the micelle. Thus, a few fast experiments enable the identification of helical regions and determination of the helix orientation within the micelle without the need for covalent modification, isotopic labeling, or sophisticated equipment. This approach opens a path toward the topology determination of alpha-helical membrane-proteins without the need for a complete NOE-based structure determination.
authors.affiliation: Institute of Chemistry/Organic and Bioorganic Chemistry, University of Graz, Heinrichstrasse 28, A-8010 Graz, Austria.
authors.forename: Michal
authors.initials: M
authors.lastname: Respondek
authors.forename: Tobias
authors.initials: T
authors.lastname: Madl
authors.forename: Christoph
authors.initials: C
authors.lastname: Göbl
authors.forename: Regina
authors.initials: R
authors.lastname: Golser
authors.forename: Klaus
authors.initials: K
authors.lastname: Zangger
mesh_terms: D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods*; D019786:Gadolinium DTPA / Q000737:chemistry; D008823:Micelles; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary
chemical_list: D023181:Antimicrobial Cationic Peptides; D008823:Micelles; C513873:antimicrobial hybrid peptide CM15; C064925:gadodiamide; D019786:Gadolinium DTPA