Solubilisation & purification of membrane proteins using benzylamine-modified SMA polymers

Aneel Akram, Waled Hadasha, Gestél C. Kuyler, Michael-Phillip Smith, Shauna Bailey-Dallaway, Aiden Preedy, Caolan Browne, Luke Broadbent, Adam Hill, Tahreem Javaid, Haroon Nazar, Nikita Samra, Anadil Naveed, Holly Tregunna, Hetal Joshi, Nusheen Akhtar, Aneesa Javed, Jessica Bowater, Joel Ravenhill, Patrik HajduYazdan Ali, Yanik Tailor, Sabreen Mumtaz, Mohammed Hamza, Kiran Gill, Jemma Gillett, Faye Patton, Huma Arshid, Maria Zaheer, Hannah Qureshi, Isabel Edwards, Shreya Patel, Aaminah Azadi, Naomi Pollock, Philip Kitchen, Bert Klumperman, Alice j. Rothnie

Research output: Contribution to journalArticlepeer-review

Abstract

Extraction of proteins from the membrane using styrene maleic acid co-polymers (SMA), forming SMA lipid particles (SMALPs), has allowed for the first time the purification of membrane proteins with their lipid bilayer environment. To date, SMA2000 has been the most effective polymer used for this purpose, with a 2:1 ratio of styrene:maleic acid, and styrene and maleic acid moieties spread statistically throughout the chain. However, SMA2000 is a highly polydisperse polymer that contains an array of different polymer lengths and sequences. RAFT polymerisation offers much better control over the polymer length; however, homogeneous distribution of styrene and maleic acid throughout the polymer is difficult to achieve. Instead, here RAFT polymerisation was used to produce a 1:1 styrene:maleic anhydride polymer, which was then modified with benzylamine. This mimics the 2:1 hydrophobic:hydrophilic nature of SMA2000, while controlling the length and obtaining a homogeneous distribution of the hydrophobic moieties (styrene and N-benzylmaleimide). SMA-benzylamine (SMA-BA) polymers of three different lengths (2, 4, and 7 kDa) were all able to solubilise purified lipids, cellular membranes, and a range of specific proteins. However, the larger 7 kDa polymer solubilised membranes more slowly and less efficiently than the shorter polymers. This also affected the yield of purified protein obtained by affinity purification with this polymer. The smallest 2 kDa polymer solubilised membranes the fastest but appeared to offer less stability to the extracted proteins. The SMA-BA polymers were more sensitive to Mg2+ ions than SMA2000. SMA-BA 4 kDa was otherwise comparable to SMA2000 and even gave a higher degree of purity.
Original languageEnglish
Article number107343
Number of pages11
JournalBiophysical Chemistry
Volume316
Early online date18 Oct 2024
DOIs
Publication statusPublished - Jan 2025

Bibliographical note

Copyright © 2024, The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/).

Data Access Statement

The underlying data for this study can be found at the Aston Data Explorer repository (https://doi.org/10.17036/researchdata.aston.ac.uk.00000647)

Keywords

  • Atm1
  • Detergent-free
  • Mass photometry
  • Nanoparticle
  • SMALP

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  • Data underpinning 'Solubilisation & purification of membrane proteins using benzylamine-modified SMA polymers

    Akram, A. (Creator), Hadasha, W. (Creator), Kuyler, G. C. (Creator), Smith, M.-P. (Creator), Bailey-Dallaway, S. (Creator), Preedy, A. (Creator), Browne, C. (Creator), Broadbent, L. (Creator), Hill, A. (Creator), Javaid, T. (Creator), Nazar, H. (Creator), Samra, N. (Creator), Naveed, A. (Creator), Tregunna, H. (Creator), Joshi, H. (Creator), Akhtar, N. (Creator), Javed, A. (Creator), Bowater, J. (Creator), Ravenhill, J. (Creator), Hajdu, P. (Creator), Ali, Y. (Creator), Tailor, Y. (Creator), Mumtaz, S. (Creator), Hamza, M. (Creator), Gill, K. (Creator), Gillett, J. (Creator), Patton, F. (Creator), Arshid, H. (Creator), Zaheer, M. (Creator), Qureshi, H. (Creator), Edwards, I. (Creator), Patel, S. (Creator), Azadi, A. (Creator), Pollock, N. (Creator), Kitchen, P. (Creator), Klumperman, B. (Creator) & Rothnie, A. (Creator), Aston Data Explorer, 4 Oct 2024

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