A Coiled-Coil Peptide Shaping Lipid Bilayers upon Fusion.

ABSTRACT: A system based on two designed peptides, namely the cationic peptide K, (KIAALKE)₃, and its complementary anionic counterpart called peptide E, (EIAALEK)₃, has been used as a minimal model for membrane fusion, inspired by SNARE proteins. Although the fact that docking of separate vesicle populations via the formation of a dimeric E/K coiled-coil complex can be rationalized, the reasons for the peptides promoting fusion of vesicles cannot be fully explained. Therefore it is of significant interest to determine how the peptides aid in overcoming energetic barriers during lipid rearrangements leading to fusion. In this study, investigations of the peptides' interactions with neutral PC/PE/cholesterol membranes by fluorescence spectroscopy show that tryptophan-labeled K? binds to the membrane (K_K? ?6.2 10³ M^-1), whereas E? remains fully water-solvated. ¹⁵N-NMR spectroscopy, depth-dependent fluorescence quenching, CD-spectroscopy experiments, and MD simulations indicate a helix orientation of K? parallel to the membrane surface. Solid-state ³¹P-NMR of oriented lipid membranes was used to study the impact of peptide incorporation on lipid headgroup alignment. The membrane-immersed K? is found to locally alter the bilayer curvature, accompanied by a change of headgroup orientation relative to the membrane normal and of the lipid composition in the vicinity of the bound peptide. The NMR results were supported by molecular dynamics simulations, which showed that K reorganizes the membrane composition in its vicinity, induces positive membrane curvature, and enhances the lipid tail protrusion probability. These effects are known to be fusion relevant. The combined results support the hypothesis for a twofold role of K in the mechanism of membrane fusion: 1) to bring opposing membranes into close proximity via coiled-coil formation and 2) to destabilize both membranes thereby promoting fusion.

SUBMITTER: Rabe M 

PROVIDER: S-EPMC5113151 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature