Membrane proteins are important pharmaceutical targets that naturally exist in complex lipid bilayer environments. As studies have increasingly shown that membrane protein structure and function are sensitive to environment, new techniques are being developed to solubilize purified membrane proteins that better replicate the complexity of a membrane.  Nanodiscs are a solubilization technique that contain a lipid bilayer 9-13 Å in diameter encapsulated by an amphipathic scaffold protein. The lipid composition of a nanodisc can be varied to include diverse phospholipids, sphingolipids, and cholesterol. However, due to the constraint of the scaffold protein, physical properties of the bilayer in a nanodisc may be different than those in a cellular bilayer even if the lipid composition is the same. The aim of this study is to examine the fluidity of lipids in nanodiscs synthesized with varying lipid compositions and compare those fluidity measurements to known values for cellular membranes. Fluidity measurements will be obtained using a fluorescence based Laurdan assay. Our preliminary data shows that we have successfully expressed and purified the MSP1D1 scaffold protein, a vital step in the synthesis of nanodiscs. This presentation will focus on optimization of diverse lipid nanodisc synthesis and baseline fluidity data from control nanodiscs of homogenous lipid compositions.

Evaluating the fluidity of diverse lipid nanodiscs

ACS Spring 2022

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