Synthesis and Evaluation of a Library of Alternating Amphipathic Copolymers to Solubilize and Study Membrane Proteins
Kopf, Adrian H.; Lijding, Odette; Elenbaas, Barend O. W.; Koorengevel, Martijn C.; Dobruchowska, Justyna M.; Walree, Cornelis A. van; Killian, J. Antoinette
(2022) Biomacromolecules, volume 23, issue 3, pp. 743 - 759
(Article)
Abstract
Amphipathic copolymers such as poly(styrene-maleic acid) (SMA) are promising tools for the facile extraction of membrane proteins (MPs) into native nanodiscs. Here, we designed and synthesized a library of well-defined alternating copolymers of SMA analogues in order to elucidate polymer properties that are important for MP solubilization and stability. MP
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extraction efficiency was determined using KcsA from E. coli membranes, and general solubilization efficiency was investigated via turbidimetry experiments on membranes of E. coli, yeast mitochondria, and synthetic lipids. Remarkably, halogenation of SMA copolymers dramatically improved solubilization efficiency in all systems, while substituents on the copolymer backbone improved resistance to Ca2+. Relevant polymer properties were found to include hydrophobic balance, size and positioning of substituents, rigidity, and electronic effects. The library thus contributes to the rational design of copolymers for the study of MPs.
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Keywords: Bioengineering, Biomaterials, Polymers and Plastics, Materials Chemistry
ISSN: 1525-7797
Publisher: American Chemical Society : Division of Carbohydrate Chemistry
Note: Funding Information: We are grateful to Anton I. P. M. de Kroon for assistance with editing parts of the manuscript, Helene Jahn for preparing the yeast mitochondrial membranes, and Bonny W. M. Kuipers for support in DLS analysis as well as Polyscope Polymers (NL) for gifting the SMAnh(2:1) copolymer (Xiran SZ30010). This work used the EM facilities at the Grenoble Instruct-ERIC Center (ISBG; UAR 3518 CNRS CEA-UGA-EMBL) with support from the French Infrastructure for Integrated Structural Biology (FRISBI; ANR-10-INSB-05-02) and GRAL, a project of the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE-0003) within the Grenoble Partnership for Structural Biology. The IBS Electron Microscope facility is supported by the Auvergne Rhône-Alpes Region, the Fonds Feder, the Fondation pour la Recherche Médicale, and GIS-IBiSA. We thank Daphna Fenel, Christine Moriscot, and Guy Schoehn, all from the Electron Microscopy platform of the Institut de Biologie Structurale, CEA, CNRS, Université Grenoble Alpes Grenoble France for performing the TEM imaging. This work was supported financially by the Division of Chemical Sciences (CW) of the Dutch Research Council (Nederlandse Organisatie voor Wetenschappelijk Onderzoek, NWO) via ECHO grant No. 711-017-006 (A.H.K.). Funding Information: We are grateful to Anton I. P. M. de Kroon for assistance with editing parts of the manuscript, Helene Jahn for preparing the yeast mitochondrial membranes, and Bonny W. M. Kuipers for support in DLS analysis as well as Polyscope Polymers (NL) for gifting the SMAnh(2:1) copolymer (Xiran SZ30010). This work used the EM facilities at the Grenoble Instruct-ERIC Center (ISBG; UAR 3518 CNRS CEA-UGA-EMBL) with support from the French Infrastructure for Integrated Structural Biology (FRISBI; ANR-10-INSB-05-02) and GRAL, a project of the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE-0003) within the Grenoble Partnership for Structural Biology. The IBS Electron Microscope facility is supported by the Auvergne Rh?ne-Alpes Region, the Fonds Feder, the Fondation pour la Recherche M?dicale, and GIS-IBiSA. We thank Daphna Fenel, Christine Moriscot, and Guy Schoehn, all from the Electron Microscopy platform of the Institut de Biologie Structurale, CEA CNRS, Universit? Grenoble Alpes Grenoble France for performing the TEM imaging. This work was supported financially by the Division of Chemical Sciences (CW) of the Dutch Research Council (Nederlandse Organisatie voor Wetenschappelijk Onderzoek, NWO) via ECHO grant No. 711-017-006 (A.H.K.). Publisher Copyright: © 2022 American Chemical Society. All rights reserved.
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