We have applied rationally designed peptide anchors to a lipid bilayer interface in order to enhance the viability of a supported lipid membrane anchored via polymer tethers. Our aim is to design these supramolecular complexes to enhance their stability and biological feasibility for the study of membrane protein and their interactions. We employ alpha-helical peptides (K3L3A12K3) as anchoring molecules, where conjugation of the peptide with N-hydroxysuccinimide (NHS)-esters through amine-based coupling allows one to access a variety of chemistries for orthogonal modification. NHS-Rhodamine and NHS-Fluorescein conjugated peptides are synthesized for peptide localization within supported membranes, and NHS-PEO4-Biotin is synthesized as a biomolecular linker. Initial work shows that peptides can be effectively partitioned within surfactant micelles, which function as in vitro models for interactions with the membrane. The secondary structure of peptide was characterized with circular dichroism in order to investigate the stability of the labeled peptide with the NHS-conjugate chemistries. Lateral fluidity of the fluorescently tagged peptide was analyzed via fluorescence imaging microscopy (Confocal Microscopy) and quantified using fluorescence recovery after photobleaching (FRAP) techniques. This data is compared with the case of lipids in untethered membranes. Furthermore, we will inspect the peptide-based anchor in detail via Nuclear Magnetic Resonance (NMR), which will supply data to evaluate the structural properties of the anchor within the bilayer. Lastly, variations in the peptide sequence allow us to rationally investigate the influence of sequence on peptide anchor stability.

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