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 Hajdu; Yazdan 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

doi:10.1016/j.bpc.2024.107343

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 journal › Article › peer-review

6 Citations (SciVal)

38 Downloads (Pure)

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 language	English
Article number	107343
Number of pages	11
Journal	Biophysical Chemistry
Volume	316
Early online date	18 Oct 2024
DOIs	https://doi.org/10.1016/j.bpc.2024.107343
Publication status	Published - Jan 2025

Bibliographical note

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)

Funding

A Akram was the recipient of a Health and Life Sciences School Studentship. SBD and CB are part of the MIBTP (Midlands Integrative Biosciences Training Partnership) doctoral training centre funded by the BBSRC (Biotechnology and Biological Sciences Research Council, BB/T00746X/1). PK is supported by the BBSRC via a Discovery Fellowship (BB/W00934X/1). BK acknowledges support through a Wellcome Trust Technology Development grant (223728/Z/21/Z). The Aston Institute for Membrane Excellence (AIME) is funded by UKRI’s Research England as part of their Expanding Excellence in England (E3) fund.

Funders	Funder number
UK Research and Innovation
Biotechnology and Biological Sciences Research Council	BB/T00746X/1, BB/W00934X/1
Wellcome Trust	223728/Z/21/Z

Keywords

Atm1
Detergent-free
Mass photometry
Nanoparticle
SMALP

Access to Document

10.1016/j.bpc.2024.107343Licence: CC BY 4.0

A Akram et al_Solubilisation and purification of membrane proteins using benzylamine-modified SMA polymers
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/).
Final published version, 3.6 MBLicence: CC BY 4.0

Cite this

Akram, A., Hadasha, W., Kuyler, G. C., Smith, M.-P., Bailey-Dallaway, S., Preedy, A., Browne, C., Broadbent, L., Hill, A., Javaid, T., Nazar, H., Samra, N., Naveed, A., Tregunna, H., Joshi, H., Akhtar, N., Javed, A., Bowater, J., Ravenhill, J., ... Rothnie, A. J. (2025). Solubilisation & purification of membrane proteins using benzylamine-modified SMA polymers. Biophysical Chemistry, 316, Article 107343. https://doi.org/10.1016/j.bpc.2024.107343

@article{7182a0380c5b4df4bd54705bc45f6c9e,

title = "Solubilisation & purification of membrane proteins using benzylamine-modified SMA polymers",

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.",

keywords = "Atm1, Detergent-free, Mass photometry, Nanoparticle, SMALP",

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

note = "Copyright {\textcopyright} 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/).",

year = "2025",

month = jan,

doi = "10.1016/j.bpc.2024.107343",

language = "English",

volume = "316",

journal = "Biophysical Chemistry",

issn = "0301-4622",

publisher = "Elsevier",

}

Akram, A, Hadasha, W, Kuyler, GC, Smith, M-P, Bailey-Dallaway, S, Preedy, A, Browne, C, Broadbent, L, Hill, A, Javaid, T, Nazar, H, Samra, N, Naveed, A, Tregunna, H, Joshi, H, Akhtar, N, Javed, A, Bowater, J, Ravenhill, J, Hajdu, P, Ali, Y, Tailor, Y, Mumtaz, S, Hamza, M, Gill, K, Gillett, J, Patton, F, Arshid, H, Zaheer, M, Qureshi, H, Edwards, I, Patel, S, Azadi, A, Pollock, N , Kitchen, P, Klumperman, B & Rothnie, AJ 2025, 'Solubilisation & purification of membrane proteins using benzylamine-modified SMA polymers', Biophysical Chemistry, vol. 316, 107343. https://doi.org/10.1016/j.bpc.2024.107343

TY - JOUR

T1 - Solubilisation & purification of membrane proteins using benzylamine-modified SMA polymers

AU - Akram, Aneel

AU - Hadasha, Waled

AU - Kuyler, Gestél C.

AU - Smith, Michael-Phillip

AU - Bailey-Dallaway, Shauna

AU - Preedy, Aiden

AU - Browne, Caolan

AU - Broadbent, Luke

AU - Hill, Adam

AU - Javaid, Tahreem

AU - Nazar, Haroon

AU - Samra, Nikita

AU - Naveed, Anadil

AU - Tregunna, Holly

AU - Joshi, Hetal

AU - Akhtar, Nusheen

AU - Javed, Aneesa

AU - Bowater, Jessica

AU - Ravenhill, Joel

AU - Hajdu, Patrik

AU - Ali, Yazdan

AU - Tailor, Yanik

AU - Mumtaz, Sabreen

AU - Hamza, Mohammed

AU - Gill, Kiran

AU - Gillett, Jemma

AU - Patton, Faye

AU - Arshid, Huma

AU - Zaheer, Maria

AU - Qureshi, Hannah

AU - Edwards, Isabel

AU - Patel, Shreya

AU - Azadi, Aaminah

AU - Pollock, Naomi

AU - Kitchen, Philip

AU - Klumperman, Bert

AU - Rothnie, Alice j.

PY - 2025/1

Y1 - 2025/1

N2 - 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.

AB - 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.

KW - Atm1

KW - Detergent-free

KW - Mass photometry

KW - Nanoparticle

KW - SMALP

UR - https://linkinghub.elsevier.com/retrieve/pii/S0301462224001728

UR - http://www.scopus.com/inward/record.url?scp=85206875599&partnerID=8YFLogxK

U2 - 10.1016/j.bpc.2024.107343

DO - 10.1016/j.bpc.2024.107343

M3 - Article

SN - 0301-4622

VL - 316

JO - Biophysical Chemistry

JF - Biophysical Chemistry

M1 - 107343

ER -

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

Abstract

Bibliographical note

Data Access Statement

Funding

Keywords

Access to Document

Other files and links

Fingerprint

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

Cite this

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

Abstract

Bibliographical note

Data Access Statement

Funding

Keywords

Access to Document

Other files and links

Fingerprint

Datasets

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

Cite this