Ciara Frances Pugh, Lukas P. Feilen, Dušan Živković, Kaia Fiil Præstegaard, Charalampos Sideris, Neil Jean Borthwick, Casper de Lichtenberg, Jani Reddy Bolla, Anton A. A. Autzen, Henriette Elisabeth Autzen
Amphiphilic copolymers capable of extracting membrane proteins directly from cellular membranes into “native nanodiscs” offer a simplified approach for preparing membrane proteins in lipid nanodiscs compared to approaches that rely on detergent. Copolymer amphiphilicity, length, and composition influence their performance, in addition to the protein itself and the purification conditions used. Here, we report a copolymer composed of methacrylic acid and styrene, which we term MAASTY, leveraging the inherent monomer reactivity ratios to create an anionic copolymer with a statistical distribution of monomers. We show that MAASTY can be used for high-resolution structural determination of a human membrane protein by single particle cryo-electron microscopy, preserving endogenous lipids including cholesterol and exhibiting an enrichment of phosphatidylinositol. Moreover, MAASTY copolymers effectively solubilize a broad range of lipid species and a wide range of different, eukaryotic membrane proteins from mammalian cells. We find that MAASTY copolymers are promising as effective solubilizers of membrane proteins and offer a chemical platform for structural and functional characterization of membrane proteins in native nanodiscs. MAASTY, a methacrylic acid–styrene copolymer, enables high-resolution structural determination of human membrane proteins in native nanodiscs, efficiently solubilizing diverse membrane proteins and preserving endogenous lipids for structural and functional studies.