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Optimization Of Detergent-Mediated Reconstitution Of Influenza A M2 Protein Into Proteoliposomes, Catherine Hirshfeld Crouch, Margaret H. Bost , '17, Tae H. Kim , '14, Bryan M. Green , '16, Douglas Stuart Arbuckle , '17, Carl H. Grossman, Kathleen P. Howard
Optimization Of Detergent-Mediated Reconstitution Of Influenza A M2 Protein Into Proteoliposomes, Catherine Hirshfeld Crouch, Margaret H. Bost , '17, Tae H. Kim , '14, Bryan M. Green , '16, Douglas Stuart Arbuckle , '17, Carl H. Grossman, Kathleen P. Howard
Physics & Astronomy Faculty Works
We report the optimization of detergent-mediated reconstitution of an integral membrane-bound protein, full-length influenza M2 protein, by direct insertion into detergent-saturated liposomes. Detergent-mediated reconstitution is an important method for preparing proteoliposomes for studying membrane proteins, and must be optimized for each combination of protein and membrane constituents used. The purpose of the reconstitution was to prepare samples for site-directed spin-labeling electron paramagnetic resonance (SDSL-EPR) studies. Our goals in optimizing the protocol were to minimize the amount of detergent used, reduce overall proteoliposome preparation time, and confirm the removal of all detergent. The liposomes were comprised of (1-palmitoyl-2-oleyl-sn-glycero-phosphocholine (POPC) and 1-palmitoyl-2-oleyl-sn-glycero-3-[phospho-rac-(1-glycerol)] …