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Tailoring butyl methacrylate/methacrylic acid copolymers for the solubilization of membrane proteins: the influence of composition and molecular weight
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SYSNO ASEP 0562629 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Tailoring butyl methacrylate/methacrylic acid copolymers for the solubilization of membrane proteins: the influence of composition and molecular weight Author(s) Janata, Miroslav (UMCH-V) RID
Čadová, Eva (UMCH-V) RID
Angelisová, Pavla (UMG-J) RID
Charnavets, Tatsiana (BTO-N)
Hořejší, Václav (UMG-J) RID
Raus, Vladimír (UMCH-V) RID, ORCIDArticle number 2200284 Source Title Macromolecular Bioscience. - : Wiley - ISSN 1616-5187
Roč. 22, č. 10 (2022)Number of pages 7 s. Language eng - English Country DE - Germany Keywords amphiphilic copolymers ; isolation ; membrane proteins Subject RIV CD - Macromolecular Chemistry OECD category Polymer science Subject RIV - cooperation Institute of Molecular Genetics - Genetics ; Molecular Biology
Institute of Biotechnology - Genetics ; Molecular BiologyR&D Projects GA19-04047S GA ČR - Czech Science Foundation (CSF) LM2018127 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Research Infrastructure CIISB II - 90127 - Masarykova univerzita Method of publishing Limited access Institutional support UMCH-V - RVO:61389013 ; UMG-J - RVO:68378050 ; BTO-N - RVO:86652036 UT WOS 000842344300001 EID SCOPUS 85136506500 DOI 10.1002/mabi.202200284 Annotation Low-molecular weight (MW) amphiphilic copolymers have been recently introduced as a powerful tool for the detergent-free isolation of cell membrane proteins. Herein, a screening approach is used to identify a new copolymer type for this application. Via a two-step ATRP/acidolysis procedure, a 3 × 3 matrix of well-defined poly[(butyl methacrylate)-co-(methacrylic acid)] copolymers (denoted BMAA) differing in their MW and ratio of hydrophobic (BMA) and hydrophilic (MAA) units is prepared. Subsequently, using the biologically relevant model (T-cell line Jurkat), two compositions of BMAA copolymers are identified that solubilize cell membranes to an extent comparable to the industry standard, styrene-maleic acid copolymer (SMA), while avoiding the potentially problematic phenyl groups. Surprisingly, while only the lowest-MW variant of the BMA/MAA 2:1 composition is effective, all the copolymers of the BMA/MAA 1:1 composition are found to solubilize the model membranes, including the high-MW variant (MW of 14 000). Importantly, the density gradient ultracentrifugation/sodium dodecyl sulfate-polyacrylamide gel electrophoresis/Western blotting experiments reveal that the BMA/MAA 1:1 copolymers disintegrate the Jurkat membranes differently than SMA, as demonstrated by the different distribution patterns of two tested membrane protein markers. This makes the BMAA copolymers a useful tool for studies on membrane microdomains differing in their composition and resistance to membrane-disintegrating polymers. Workplace Institute of Macromolecular Chemistry Contact Eva Čechová, cechova@imc.cas.cz ; Tel.: 296 809 358 Year of Publishing 2023 Electronic address https://onlinelibrary.wiley.com/doi/10.1002/mabi.202200284
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