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

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 pu...

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Bibliographic Details
Published in:	Biophysical chemistry Vol. 316; p. 107343
Main Authors:	Akram, Aneel, Hadasha, Waled, Kuyler, Gestél C., Smith, Michael-Phillip, Bailey-Dallaway, Shauna, Preedy, Aiden, Browne, Caolan, Broadbent, Luke, Hill, Adam, Javaid, Tahreem, Nazar, Haroon, Samra, Nikita, Naveed, Anadil, Tregunna, Holly, Joshi, Hetal, Akhtar, Nusheen, Javed, Aneesa, Bowater, Jessica, Ravenhill, Joel, Hajdu, Patrik, Ali, Yazdan, Tailor, Yanik, Mumtaz, Sabreen, Hamza, Mohammed, Gill, Kiran, Gillett, Jemma, Patton, Faye, Arshid, Huma, Zaheer, Maria, Qureshi, Hannah, Edwards, Isabel, Patel, Shreya, Azadi, Aaminah, Pollock, Naomi, Kitchen, Philip, Klumperman, Bert, Rothnie, Alice J.
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 01-01-2025
Subjects:	Atm1 Detergent-free Mass photometry Nanoparticle SMALP Mn Nanoparticle CD81 BPT E.coli AQP4 DLS ZipA Ð BSA DIBMA THF PDI SMAnh FTIR SMA-BA 7 kDa RAFT DMF DMPC SMALP SMA1000-BA GSSG SMA Detergent-free Sf9 SMA-BA 2 kDa Atm1 AIBN MAnh MRP4 P.pastoris SEC SDS-PAGE MEK SMA-BA 4 kDa LB Mass photometry ABC transporter Ag-GSH Ni-NTA ATR BA
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Summary:	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. [Display omitted] •1:1 styrene:maleic anhydride polymers were generated by RAFT polymerisation and modified with benzylamine.•All SMA-benzylamine (SMA-BA) polymers were able to solubilise a range of proteins from different expression systems.•Shorter polymers solubilised membranes more quickly than longer polymers, but the formed particles were less stable.•SMA-BA polymers are more sensitive to divalent cations than SMA2000.•SMA-BA 4 kDa was comparable to SMA2000 for solubilisation efficiency, purification yield, and protein ligand binding.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0301-4622 1873-4200 1873-4200
DOI:	10.1016/j.bpc.2024.107343