(228aa) Characterization of Molecularly-Complete Planar Supported Cell Plasma Membranes for Biomedical Applications

Emerging technologies to study membrane proteins and protein-lipid interactions use cell plasma membrane blebs as an intermediate to form planar bilayers that are compatible with a variety of characterization tools and microscopy methods. This technology allows membrane proteins to be incorporated into supported lipid bilayers without using detergents and reconstitution, preserving the native lipids and other species within the plasma membrane. However, the impact of chemicals used to induce cell blebbing on protein properties is still unknown. Our research focuses on characterization of the cell blebs under various blebbing conditions and the result on protein behavior (mobility, activity, orientation, etc.) in this emerging platform. Bilayers formed from cell blebs expressing GPI-linked yellow fluorescent protein (GPI-YFP) provide a simple way for us to study the mobility of proteins through the single particle tracking (SPT) and how properties change, e.g., confinement effects, immobility, etc., with the conditions used for bleb generation. The orientation of GPI-YFP in the cell blebs and bleb bilayer is revealed using a protease cleavage assay. We will discuss these characterizations of supported cell plasma membranes and the application of this platform to create adult stem cell bilayers for the study of oncogenic microvesicle interactions.