Membrane proteins were studied using detergent for extraction, which was an expensive and difficult technique. But now Styrene Maleic anhydride copolymers (SMA) are being used for providing native like environment to study the characteristics and structure of membrane proteins. Challenge with this technique is the styrene ring and protein both absorb UV-Vis at 280nm. In this study, styrene ring was replaced with Dodecyl vinyl ether (DVE) and Butyl vinyl ether (BVE). Optimization of the ratio of maleic anhydride (MAn), BVE and DVE copolymer was synthesized using reversible addition−fragmentation chain-transfer (RAFT) technique. Endgroup of copolymer was removed and hydrolysis was performed on it. A thin layer of 3:1 POPC and POPG was used to interact with lipid. Optimized ratio and a few other ratios were tested against lipids. DLS and TEM images show formation of lipid discs. It was observed that freeze thaw and shaking overnight help in making disc size smaller and more uniform. Initially, it was observed that disc size was affected by changing mole ratio, increasing number of freeze thaw cycles for penetration of polymers and agitating overnight. This study will further focus on comparison of different mole ratios of these monomers and choosing one optimized hydrophobic monomer instead of two vinyl ethers.

Lipid discs formation for studying membrane proteins

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