Developing Glycolipid Biointerfaces to Decipher Ganglioside-EGFR Lateral Interactions

The dense layer of glycans coating of the exterior of cell membranes, known as the glycocalyx, mediates a broad range of cellular processes. Numerous studies have indicated the importance of glycocalyx composition on disease pathology ranging from insulin resistance to cancer proliferation. However, due to the incredible complexity of cell-surface glycan composition, the identification of the key components mediating particular lipid-protein interactions has had very limited success. In this study we present our current work on developing novel cell-surface mimicking biointerfaces that allow for study of the impacts of membrane lipid components on membrane protein function and signaling. Combining this interface with the surface plasmon resonance (SPR) sensing modality, we investigate the impact of membrane ganglioside composition on the binding behavior of the transmembrane oncoprotein epidermal growth factor receptor (EGFR) as a model system. Through induced vesiculation and capture of cell membranes containing EGFR, biomimetic sensing interfaces were formed on SPR substrates where the binding behavior of EGFR can be tested in a highly sensitive and label-free manner. Furthermore, a diverse ganglioside library can be introduced into these membranes, thus allowing for investigation and quantification of their individual effects on EGFR’s binding activity. Here we introduce our work on the formation and characterization of the biomimetic platform, with preliminary results indicating the value of this approach for the study of a wide range of traditionally difficult to elucidate membrane protein interactions.