Biophys J, November 2000, p. 2644-2656, Vol. 79, No. 5

The Topology of Lysine-Containing Amphipathic Peptides in Bilayers by Circular Dichroism, Solid-State NMR, and Molecular Modeling

Bas Vogt,^* Philippe Ducarme, Susan Schinzel,^* Robert Brasseur, and Burkhard Bechinger^*

 ^*Max-Planck-Institut für Biochemie, 82152 Martinsried, Germany; and  Centre de Biophysique Moléculaire Numérique, Faculté Universitaire des Sciences Agronomique de Gembloux, Passage des Déportés, 5030 Gembloux, Belgium

In order to better understand the driving forces that determine the alignment of amphipathic helical polypeptides with respect to the surface of phospholipid bilayers, lysine-containing peptide sequences were designed, prepared by solid-phase chemical synthesis, and reconstituted into membranes. CD spectroscopy indicates that all peptides exhibit a high degree of helicity in the presence of SDS micelles or POPC small unilamellar vesicles. Proton-decoupled ³¹P-NMR solid-state NMR spectroscopy demonstrates that in the presence of peptides liquid crystalline phosphatidylcholine membranes orient well along glass surfaces. The orientational distribution and dynamics of peptides labeled with ¹⁵N at selected sites were investigated by proton-decoupled ¹⁵N solid-state NMR spectroscopy. Polypeptides with a single lysine residue adopt a transmembrane orientation, thereby locating this polar amino acid within the core region of the bilayer. In contrast, peptides with 3 lysines reside along the surface of the membrane. With 2 lysines in the center of an otherwise hydrophobic amino acid sequence the peptides assume a broad orientational distribution. The energy of lysine discharge, hydrophobic, polar, and all other interactions are estimated to quantitatively describe the polypeptide topologies observed. Furthermore, a molecular modeling algorithm based on the hydrophobicities of atoms in a continuous hydrophilic-hydrophobic-hydrophilic potential describes the experimentally observed peptide topologies well.

Biophys J, November 2000, p. 2644-2656, Vol. 79, No. 5
© 2000 by the Biophysical Society   0006-3495/00/11/2644/13  $2.00

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