Tether-Supported Biomembranes with α‑Helical Peptide-Based Anchoring Constructs

• Published in: Langmuir Vol. 29; no. 1; pp. 299 - 307
• Main Authors: Zhong, Lina, Tu, Raymond, Gilchrist, M. Lane
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 08.01.2013
• Subjects: Biological and medical sciences; Biological membranes; Chemistry; Chromatography, High Pressure Liquid; Circular Dichroism; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; General and physical chemistry; Lipids - chemistry; Membrane physicochemistry; Membrane Proteins - chemistry; Models, Biological; Molecular biophysics; Peptides - chemistry; Polyethylene Glycols - chemistry; Silicon Dioxide - chemistry; Solid-Phase Synthesis Techniques; Surface Properties; Biomembrane; Peptides; Support; Anchoring
• ISSN: 0743-7463; 1520-5827
• DOI: 10.1021/la303628n

The strict requirement of constructing a native lipid environment to preserve the structure and functionality of membrane proteins is the starting constraint when building biomaterials and sensor systems from these biomolecules. To enhance the viability of supported biomembranes systems and build new ligand display interfaces, we apply rationally designed peptides partitioned into the lipid bilayer interface. Peptides designed to form membrane-spanning α-helical anchoring domains are synthesized using solid-phase peptide synthesis. K3A4L2A7L2A3K2-FITC is synthesized on the 100 mg scale for use as a biomembrane anchoring molecule, where orthogonal side-chain modifications allow us to introduce probes enabling peptide localization within supported bilayers. The peptides are found to form α-helical domains within liposomes as assessed with circular dichroism spectroscopy. These peptides are designed to be incorporated into lipid bilayers supported by microspheres and serve as biomembrane anchoring moieties to amino-terminated surfaces. Here, the silica bead surface (4.7 μm diameter) is activated with homobifunctional NHS-PEG3000-NHS as “polymer cushion” spacers. This tethering to a subset of the K3A4L2A7L2A3K2-FITC molecules present in the bilayer is achieved by the fusion of liposomes followed by coupling of the peptide amino groups to the NHS presented from the silica microsphere surfaces. The biomembrane distributions of tethered and untethered K3A4L2A7L2A3K2-FITC are probed with confocal microscopy and are found to give 3D reconstructions consistent with largely homogeneous supported biomembranes. The fluidity of the untethered fraction of peptides within supported membranes is quantified using the fluorescence recovery after photobleaching (FRAP) technique. The presence of the PEG3000 polymer cushion facilitated a 28.9% increase in peptide diffusivity over untethered bilayers at the lowest peptide to lipid ratio we examined. We show that rationally designed peptide-based anchors can be used to tether lipid bilayers, creating a polymer-cushioned lipid microenvironment on surfaces with high lateral mobility and facilitating the development of a new platform for ligand displays.