Recent Advances on Polymer Lipid Particles (PoLP) in Membrane Protein Research

Abstract

Membrane proteins (MPs) structure elucidation is of crucial importance as they represent a major target for drug design. However, owing to their high hydrophobicity, MPs are challenging proteins to study. The use of polymer lipid nanoparticles (PoLP) has recently allowed a leap forward in the field of MP research. A wide range of polymers have been successfully used for elucidating the structure of MPs. One polymer in particular, styrene maleic acid (SMA) copolymer, is recently catching a large interest owing to its easy use in detergentā€free membrane solubilisation and the recent successes in purification and stabilising MPs for further biophysical studies.

Key Concepts

  • SMA copolymers are new tools to purify membrane proteins.
  • SMA copolymers can solubilise membrane proteins directly without the need of detergents.
  • Membrane proteins purified in SMALP are stable over time and remain active owing to the retention of native lipids around the proteins.
  • SMA use limitations include acid pH sensitivity, divalent cations sensitivity and UV absorption.
  • New polymers (SMA and others) are being engineered to overcome these limitations.

Keywords: membrane protein; polymer; styrene; maleic acid; SMA; nanoparticles; PoLP

Figure 1. Schematic of the polymerisation and alkaline hydrolysis events leading to the formation of the styrene maleimide copolymer 2:1.
Figure 2. Solubilisation stages and nanoparticle generation. (a) Representation of a membrane protein in a SMA nanoparticle. (b) Schematic of the putative steps involved in the solubilisation of membranes by SMA copolymer to form nanoparticles.
Figure 3. Purification of membrane protein with SMA copolymer solubilisation. Schematic explaining the various steps of SMA solubilisation of membranes, nanoparticle generation, purifcation via a His‐tag (red star) on the protein of interest and further particle analysis.
close

References

Arunmanee W, Harris JR and Lakey JH (2014) Outer membrane protein F stabilised with minimal amphipol forms linear arrays and LPS‐dependent 2D crystals. Journal of Membrane Biology 247: 949–956.

Belkhiria S, Thierry M and Albert R (1994) Styrene maleic anhydride copolymerization in a recycle tubular reactor: reactor stability and product quality. Chemical Engineering Science 49: 4981–4990.

Bell AJ, Frankel LK and Bricker TM (2015) High yield non‐detergent isolation of photosystem I‐light‐harvesting chlorophyll II membranes from spinach thylakoids: implications for the organisation of the PS I antennae in higher plants. Journal of Biological Chemistry 290: 18429–18437.

Broecker J, Eger BT and Ernst OP (2017) Crystallogenesis of membrane proteins mediated by polymer‐bounded lipid nanodiscs. Structure 25: 384–392.

Calabrese AN, Watkinson TG, Henderson PJF, Radford SE and Ashcroft AE (2015) Amphipols outperform dodecylmaltoside micelles in stabilizing membrane protein structure in the gas phase. Analytical Chemistry 87: 1118–1126.

Chawla U, Jiang Y, Zheng W, et al. (2016) A usual G‐protein‐coupled receptor in unusual membranes. Angewandte Chemie International Edition in English 55: 588–592.

Craig AF, Clark EE, Sahu ID, et al. (2016) Tuning the size of styrene–maleic acid copolymer–lipid nanoparticles (SMALPs) using RAFT polymerization for biophysical studies. Biochimica et Biophysica Acta 1858: 2931–2939.

Cuevas Arenas R, Danielczak B, Martel A, et al. (2017) Fast collisional lipid transfer among polymer‐bounded nanodiscs. Scientific Reports 7: 45875.

Dörr JM, Koorengevel MC, Schäfer M, et al. (2014) Detergent‐free isolation, characterization, and functional reconstitution of a tetrameric K+ channel: the power of native nanodiscs. Proceedings of the National Academy of Sciences of the United States of America 111: 18607–18612.

Dörr JM, Scheidelaar S, Koorengevel MC, et al. (2016) The styrene–maleic acid copolymer: a versatile tool in membrane research. European Biophysics Journal 45: 3–21.

Fiori MC, Jiang Y, Altenberg GA and Liang H (2017a) Polymer‐encased nanodiscs with improved buffer compatibility. Scientific Reports 7: 7432.

Fiori MC, Jiang Y, Zheng W, et al. (2017b) Polymer nanodiscs: discoidal amphiphilic block copolymer membranes as a new platform for membrane proteins. Scientific Reports 7: 15227.

Grethen A, Oluwole AO, Danielczak B, Vargas C and Keller S (2017) Thermodynamics of nanodisc formation mediated by styrene/maleic acid (2:1) copolymer. Scientific Reports 7: 11517.

Gulati S, Jamshad M, Knowles TJ, et al. (2014) Detergent‐free purification of ABC (ATP‐binding‐cassette) transporters. Biochemistry Journal 461: 269–278.

Hall SCL, Tognoloni C, Price GJ, et al. (2017) The influence of poly(styrene‐co‐maleic acid) copolymer structure on the properties and self‐assembly of SMALP nanodiscs. Biomacromolecules 19 (3): 761–772.

Jamshad M, Grimard V, Idini I, et al. (2014) Structural analysis of a nanoparticle containing a lipid bilayer used for detergent‐free extraction of membrane proteins. Nano Research 8: 774–789.

Jamshad M, Charlton J, Lin Y‐P, et al. (2015) G‐protein coupled receptor solubilization and purification for biophysical analysis and functional studies, in the total absence of detergent. Bioscience Reports 35 (2). 10.1042/BSR20140171, pii: e00188.

John Wiley & Sons, Ltd (2001) Membrane Proteins. In: Encyclopedia of Life Sciences, p. 501. Chichester, UK: John Wiley & Sons, Ltd.

Kleinschmidt JH and Popot J‐L (2014) Folding and stability of integral membrane proteins in amphipols. Archives of Biochemistry and Biophysics 564: 327–343.

Knowles TJ, Finka R, Smith C, et al. (2009) Membrane proteins solubilized intact in lipid containing nanoparticles bounded by styrene maleic acid copolymer. Journal of the American Chemical Society 131: 7484–7485.

Kumar M, Grzelakowski M, Zilles J, Clark M and Meier W (2007) Highly permeable polymeric membranes based on the incorporation of the functional water channel protein Aquaporin Z. Proceedings of the National Academy of Sciences of the United States of America 104: 20719–20724.

Lee SC, Khalid S, Pollock NL, et al. (2016a) Encapsulated membrane proteins: a simplified system for molecular simulation. Biochimica et Biophysica Acta 1858: 2549–2557.

Lee SC, Knowles TJ, Postis VLG, et al. (2016b) A method for detergent‐free isolation of membrane proteins in their local lipid environment. Nature Protocols 11: 1149–1162.

Lewis FM, Walling C, Cummings W, Briggs ER and Mayo FR (1948) Copolymerization. IV. Effects of temperature and solvents on monomer reactivity ratios. Journal of the American Chemical Society 70: 1519–1523.

Li D, Li J, Zhuang Y, et al. (2015) Nano‐size uni‐lamellar lipodisq improved in situ auto‐phosphorylation analysis of E. coli tyrosine kinase using (19)F nuclear magnetic resonance. Protein & Cell 6: 229–233.

Lindhoud S, Carvalho V, Pronk JW and Aubin‐Tam M‐E (2016) SMA‐SH: modified styrene–maleic acid copolymer for functionalization of lipid nanodiscs. Biomacromolecules 17: 1516–1522.

Logez C, Damian M, Legros C, et al. (2016) Detergent‐free isolation of functional G protein‐coupled receptors into nanometric lipid particles. Biochemistry 55: 38–48.

Long AR, O'Brien CC, Malhotra K, et al. (2013) A detergent‐free strategy for the reconstitution of active enzyme complexes from native biological membranes into nanoscale discs. BMC Biotechnology 13: 41.

Mayo FR (1943) Chain transfer in the polymerization of styrene: the reaction of solvents with free radicals. Journal of the American Chemical Society 65: 2324–2329.

Mayo FR and Lewis FM (1944) Copolymerization. I. A basis for comparing the behavior of monomers in copolymerization; the copolymerization of styrene and methyl methacrylate. Journal of the American Chemical Society 66: 1594–1601.

Mayo FR, Walling C, Lewis FM and Hulse WF (1948) Copolymerization. V.1 Some copolymerizations of vinyl acetate. Journal of the American Chemical Society 70: 1523–1525.

Morrison KA, Akram A, Mathews A, et al. (2016) Membrane protein extraction and purification using styrene–maleic acid (SMA) copolymer: effect of variations in polymer structure. Biochemistry Journal 473: 4349–4360.

Oluwole AO, Danielczak B, Meister A, et al. (2017) Solubilization of membrane proteins into functional lipid‐bilayer nanodiscs using a diisobutylene/maleic acid copolymer. Angewandte Chemie International Edition in English 56: 1919–1924.

Orwick‐Rydmark M, Lovett JE, Graziadei A, et al. (2012) Detergent‐free incorporation of a seven‐transmembrane receptor protein into nanosized bilayer lipodisq particles for functional and biophysical studies. Nano Letters 12: 4687–4692.

Parmar MJ, de Marcos Lousa C, Muench SP, Goldman A and Postis VLG (2016) Artificial membranes for membrane protein purification, functionality and structure studies. Biochemical Society Transactions 44: 877–882.

Parmar M, Rawson S, Scarff CA, et al. (2017) Using a SMALP platform to determine a sub‐nm single particle cryo‐EM membrane protein structure. Biochimica et Biophysica Acta 1860: 378–383.

Paulin S, Jamshad M, Dafforn TR, et al. (2014) Surfactant‐free purification of membrane protein complexes from bacteria: application to the staphylococcal penicillin‐binding protein complex PBP2/PBP2a. Nanotechnology 25: 285101.

Pollock NL, Lee SC, Patel JH, Gulamhussein AA and Rothnie AJ (2017) Structure and function of membrane proteins encapsulated in a polymer‐bound lipid bilayer. Biochimica et Biophysica Acta 1860: 809–817.

Polovinkin V, Gushchin I, Sintsov M, et al. (2014) High‐resolution structure of a membrane protein transferred from amphipol to a lipidic mesophase. Journal of Membrane Biology 247: 997–1004.

Popot J‐L (2014) Amphipols: where from? Where to? Journal of Membrane Biology 247: 755–757.

Prabudiansyah I, Kusters I, Caforio A and Driessen AJM (2015) Characterization of the annular lipid shell of the Sec translocon. Biochimica et Biophysica Acta 1848: 2050–2056.

Ramadugu VSK, Di Mauro GM, Ravula T and Ramamoorthy A (2017) Polymer nanodiscs and macro‐nanodiscs of a varying lipid composition. Chemical Communications 53: 10824–10826.

Ravula T, Hardin NZ, Ramadugu S, Cox SJ and Ramamoorthy A (2017a) pH resistant monodispersed polymer–lipid nanodiscs. Angewandte Chemie International Edition in English 130 (5): 1356–1359. DOI: 10.1002/ange.201712017.

Ravula T, Hardin NZ, Ramadugu SK and Ramamoorthy A (2017b) pH tunable and divalent metal ion tolerant polymer lipid nanodiscs. Langmuir. DOI: 10.1021/acs.langmuir.7b02887.

Ravula T, Ramadugu S, Di Mauro G and Ramamoorthy A (2017c) Bioinspired, size‐tunable self‐assembly of polymer–lipid bilayer nanodiscs. Angewandte Chemie International Edition in English. DOI: 10.1002/ange.201705569.

Routledge SJ, Mikaliunaite L, Patel A, et al. (2015) The synthesis of recombinant membrane proteins in yeast for structural studies. Methods 95: 26–37.

Sahu ID, McCarrick RM, Troxel KR, et al. (2013) DEER EPR measurements for membrane protein structures via bifunctional spin labels and lipodisq nanoparticles. Biochemistry 52: 6627–6632.

Sahu ID, Zhang R, Dunagan MM, Craig AF and Lorigan GA (2017) Characterization of KCNE1 inside lipodisq nanoparticles for EPR spectroscopic studies of membrane proteins. Journal of Physical Chemistry B 121: 5312–5321.

Scheidelaar S, Koorengevel MC, Pardo JD, et al. (2015) Molecular model for the solubilization of membranes into nanodisks by styrene maleic acid copolymers. Biophysical Journal 108: 279–290.

Scheidelaar S, Koorengevel MC, van Walree CA, et al. (2016) Effect of polymer composition and pH on membrane solubilization by styrene–maleic acid copolymers. Biophysical Journal 111: 1974–1986.

Schmidt V and Sturgis JN (2017) Modifying styrene‐maleic acid co‐polymer for studying lipid nanodiscs. Biochimica et Biophysica Acta 1860: 777–783.

Schulz S, Wilkes M, Mills DJ, Kühlbrandt W and Meier T (2017) Molecular architecture of the N‐type ATPase rotor ring from Burkholderia pseudomallei. EMBO Reports 18: 526–535.

Sharma KS, Durand G, Gabel F, et al. (2012) Non‐ionic amphiphilic homopolymers: synthesis, solution properties, and biochemical validation. Langmuir 28: 4625–4639.

Skaar K, Korza HJ, Tarry M, Sekyrova P and Högbom M (2015) Expression and subcellular distribution of GFP‐tagged human tetraspanin proteins in Saccharomyces cerevisiae. PLoS ONE 10: e0134041.

Smirnova IA, Sjöstrand D, Li F, et al. (2016) Isolation of yeast complex IV in native lipid nanodiscs. Biochimica et Biophysica Acta 1858: 2984–2992.

Smith AAA, Autzen HE, Laursen T, et al. (2017) Controlling styrene maleic acid lipid particles through RAFT. Biomacromolecules 18: 3706–3713.

Swainsbury DJK, Scheidelaar S, Foster N, et al. (2017) The effectiveness of styrene–maleic acid (SMA) copolymers for solubilisation of integral membrane proteins from SMA‐accessible and SMA‐resistant membranes. Biochimica et Biophysica Acta 1859: 2133–2143.

Swainsbury DJK, Scheidelaar S, van Grondelle R, Killian JA and Jones MR (2014) Bacterial reaction centers purified with styrene maleic acid copolymer retain native membrane functional properties and display enhanced stability. Angewandte Chemie International Edition in English 53: 11803–11807.

Tanaka M, Hosotani A, Tachibana Y, et al. (2015) Preparation and characterization of reconstituted lipid‐synthetic polymer discoidal particles. Langmuir 31: 12719–12726.

Tribet C, Audebert R and Popot JL (1996) Amphipols: polymers that keep membrane proteins soluble in aqueous solutions. Proceedings of the National Academy of Sciences of the United States of America 93: 15047–15050.

Vargas C, Arenas RC, Frotscher E and Keller S (2015) Nanoparticle self‐assembly in mixtures of phospholipids with styrene/maleic acid copolymers or fluorinated surfactants. Nanoscale 7: 20685–20696.

Yasuhara K, Arakida J, Ravula T, et al. (2017) Spontaneous lipid nanodisc formation by amphiphilic polymethacrylate copolymers. Journal of the American Chemical Society 139: 18657–18663.

Zhang R, Sahu ID, Liu L, et al. (2015) Characterizing the structure of lipodisq nanoparticles for membrane protein spectroscopic studies. Biochimica et Biophysica Acta 1848: 329–333.

Zhang S, Li N, Zeng W, Gao N and Yang M (2017) Cryo‐EM structures of the mammalian endo‐lysosomal TRPML1 channel elucidate the combined regulation mechanism. Protein & Cell 8: 834–847.

Zoonens M and Popot J‐L (2014) Amphipols for each season. Journal of Membrane Biology 247: 759–796.

Further Reading

Esmaili M and Overduin M (2018) Membrane biology visualized in nanometer‐sized discs formed by styrene maleic acid polymers. Biochimica et Biophysica Acta 1860 (2): 257–263.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
de Marcos Lousa, Carine, and Postis, Vincent(Jul 2018) Recent Advances on Polymer Lipid Particles (PoLP) in Membrane Protein Research. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0027944]