Abstract
Membrane proteins are involved in various critical biological processes, and studying membrane proteins represents a major challenge in protein biochemistry. As shown by both structural and functional studies, the membrane environment plays an essential role for membrane proteins. In vitro studies are reliant on the successful reconstitution of membrane proteins. This review describes the interaction between detergents and lipids that aids the understanding of the reconstitution processes. Then the techniques of detergent removal and a few useful techniques to refine the formed proteoliposomes are reviewed. Finally the applications of reconstitution techniques to study membrane proteins involved in Ca2+ signaling are summarized.
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Rigaud JL, Pitard B, Levy D. Reconstitution of membrane proteins into liposomes: application to energy-transducing membrane proteins. Biochim Biophys Acta Bioenerg, 1995, 1231: 223–246
Weinglass AB, Whitelegge JP, Kaback HR. Integrating mass spectrometry into membrane protein drug discovery. Curr Opin Drug Dis Dev, 2004, 7: 589–599
Long SB, Tao X, Campbell EB, MacKinnon R. Atomic structure of a voltage-dependent K+ channel in a lipid membrane-like environment. Nature, 2007, 450: 376–383
Gonen T, Cheng Y, Sliz P, Hiroaki Y, Fujiyoshi Y, Harrison SC, Walz T. Lipid-protein interactions in double-layered two-dimensional AQP0 crystals. Nature, 2005, 438: 633–638
Schmidt D, Jiang QX, MacKinnon R. Phospholipids and the origin of cationic gating charges in voltage sensors. Nature, 2006, 444: 775–779
Tanaka JC, Eccleston JF, Barchi RL. Cation selectivity characteristics of the reconstituted voltage-dependent sodium channel purified from rat skeletal muscle sarcolemma. J Biol Chem, 1983, 258: 7519–7526
Brohawn SG, del Mármol J, MacKinnon R. Crystal structure of the human K2P TRAAK, a lipid- and mechano-sensitive K+ ion channel. Science, 2012, 335: 436–441
Wang L, Sigworth FJ. Structure of the BK potassium channel in a lipid membrane from electron cryomicroscopy. Nature, 2009, 461: 292–295
Nakao S, Ebata H, Hamamoto T, Kagawa Y, Hirata H. Solubilization and reconstitution of voltage-dependent calcium-channel from bovine cardiac-muscle. Ca2+ influx assay using the fluorescent dye Quin2. Biochim Biophys Acta, 1988, 944: 337–343
Ramos-Franco J, Bare D, Caenepeel S, Nani A, Fill M, Mignery G. Single-channel function of recombinant type 2 inositol 1,4,5-trisphosphate receptor. Biophys J, 2000, 79: 1388–1399
Kagawa Y, Racker E. Partial resolution of the enzymes catalyzing oxidative phosphorylation: XXV. Reconstitution of vesicles catalyzing 32Pi-adenosine triphosphate exchange. J Biol Chem, 1971, 246: 5477–5487
Hinkle PC, Kim JJ, Racker E. Ion Transport and respiratory control in vesicles formed from cytochrome oxidase and phospholipids. J Biol Chem, 1972, 247: 1338–1339
Racker E. Reconstitution of a calcium pump with phospholipids and a purified Ca++-adenosine triphosphatase from sarcoplasmic reticulum. J Biol Chem, 1972, 247: 8198–8200
Shen HH, Lithgow T, Martin LL. Reconstitution of membrane proteins into model membranes: seeking better ways to retain protein activities. Int J Mol Sci, 2013, 14: 1589–1607
Ritchie TK, Grinkova YV, Bayburt TH, Denisov IG, Zolnerciks JK, Atkins WM, Sligar SG. Chapter 11 Reconstitution of membrane proteins in phospholipid bilayer nanodiscs. Methods Enzymol, 2009, 464: 211–231
Näsvik Öjemyr L, Von Ballmoos C, Gennis RB, Sligar SG, Brzezinski P. Reconstitution of respiratory oxidases in membrane nanodiscs for investigation of proton-coupled electron transfer. FEBS Lett, 2012, 586: 640–645
Shnyrova AV, Zimmerberg J. Reconstitution of membrane budding with unilamellar vesicles. Methods Enzymol, 2009, 464: 55–75
Seddon AM, Curnow P, Booth PJ. Membrane proteins, lipids and detergents: not just a soap opera. Biochim Biophys Acta Biomembr, 2004, 1666: 105–117
Pagano A, Spiess M. Reconstitution of Rab4-dependent vesicle formation in vitro. Methods Enzymol, 2005, 403: 81–92
Higgins MK, McMahon HT. In vitro reconstitution of discrete stages of dynamin-dependent endocytosis. Methods Enzymol, 2006, 404: 597–611
Kano F, Takenaka K, Murata M. Reconstitution of Golgi disassembly by mitotic Xenopus egg extract in semi-intact MDCK cells. Methods Mol Biol (Clifton, NJ), 2006, 322: 357–365
Ollivon M, Lesieur S, Grabielle-Madelmont C, Paternostre M. Vesicle reconstitution from lipid-detergent mixed micelles. Biochim Biophys Acta Biomembr, 2000, 1508: 34–50
Le Maire M, Champeil P, Møller JV. Interaction of membrane proteins and lipids with solubilizing detergents. Biochim Biophys Acta Biomembr, 2000, 1508: 86–111
Garavito RM, Ferguson-Miller S. Detergents as tools in membrane biochemistry. J Biol Chem, 2001, 276: 32403–32406
Gohon Y, Popot JL. Membrane protein-surfactant complexes. Curr Opin Colloid Interface Sci, 2003, 8: 15–22
Israelachvili JN. Intermolecular and Surface Forces. Burlington, MA: Academic Press, 2011
Lévy D, Gulik A, Bluzat A, Rigaud JL. Reconstitution of the sarcoplasmic reticulum Ca2+-ATPase: mechanisms of membrane protein insertion into liposomes during reconstitution procedures involving the use of detergents. Biochim Biophys Acta Biomembr, 1992, 1107: 283–298
Young HS, Rigaud JL, Lacapere JJ, Reddy LG, Stokes DL. How to make tubular crystals by reconstitution of detergent-solubilized Ca2+-ATPase. Biophys J, 1997, 72: 2545–2558
Seras-Cansell M, Ollivon M, Lesieur S. Generation of non-ionic monoalkyl amphiphile-cholesterol vesicles: evidence of membrane impermeability to octyl glucoside. STP Pharma Sci, 1996, 6: 12–20
Holloway PW. A simple procedure for removal of triton X-100 from protein samples. Anal Biochem, 1973, 53: 304–308
Zhou X, Graham TR. Reconstitution of phospholipid translocase activity with purified Drs2p, a type-IV P-type ATPase from budding yeast. Proc Natl Acad Sci USA, 2009, 106: 16586–16591
Kim M, Song E. Iron transport by proteoliposomes containing mitochondrial F1Fo ATP synthase isolated from rat heart. Biochimie, 2010, 92: 333–342
Schaedler TA, Tong Z, van Veen HW. The multidrug transporter LmrP protein mediates selective calcium efflux. J Biol Chem, 2012, 287: 27682–27690
Lévy D, Bluzat A, Seigneuret M, Rigaud JL. A systematic study of liposome and proteoliposome reconstitution involving Bio-Bead-mediated Triton X-100 removal. Biochim Biophys Acta Biomembr, 1990, 1025: 179–190
Mimms LT, Zampighi G, Nozaki Y, Tanford C, Reynolds JA. Phospholipid vesicle formation and transmembrane protein incorporation using octyl glucoside. Biochemistry, 1981, 20: 833–840
Kasahara M, Hinkle PC. Reconstitution and purification of the D-glucose transporter from human erythrocytes. J Biol Chem, 1977, 252: 7384–7390
Israel M, Manaranche R. The release of acetylcholine-from a cellular towards a molecular mechanism. Biol Cell, 1985, 55: 1–14
Traikia M, Warschawski DE, Recouvreur M, Cartaud J, Devaux PF. Formation of unilamellar vesicles by repetitive freeze-thaw cycles: characterization by electron microscopy and P-31-nuclear magnetic resonance. Eur Biophys J Biophys Lett, 2000, 29: 184–195
Mayer LD, Hope MJ, Cullis PR. Vesicles of variable sizes produced by a rapid extrusion procedure. Biochim Biophys Acta, 1986, 858: 161–168
Johnson SM, Bangham AD, Hill MW, Korn ED. Single bilayer liposomes. Biochim Biophys Acta Biomembr, 1971, 223: 820–826
Huang CH. Studies on phosphatidylcholine vesicles. Formation and physical characteristics. Biochemistry, 1969, 8: 344–352
Lapinski MM, Castro-Forero A, Greiner AJ, Ofoli RY, Blanchard GJ. Comparison of liposomes formed by sonication and extrusion: rotational and translational diffusion of an embedded chromophore. Langmuir, 2007, 23: 11677–11683
Bauer PJ, Drechsler M. Association of cyclic GMP-gated channels and Na+-Ca2+-K+ exchangers in bovine retinal rod outer segment plasma-membranes. J Phys Lond, 1992, 451: 109–131
Bucher K, Belli S, Wunderli-Allenspach H, Kramer SD. P-glycoprotein in proteoliposomes with low residual detergent: the effects of cholesterol. Pharm Res, 2007, 24: 1993–2004
Berridge MJ, Bootman MD, Roderick HL. Calcium signalling: dynamics, homeostasis and remodelling. Nat Rev Mol Cell Biol, 2003, 4: 517–529
Harper JF, Harmon A. Plants, symbiosis and parasites: a calcium signalling connection. Nat Rev Mol Cell Biol, 2005, 6: 555–566
Hofer AM, Brown EM. Extracellular calcium sensing and signalling. Nat Rev Mol Cell Biol, 2003, 4: 530–538
Orrenius S, Zhivotovsky B, Nicotera P. Regulation of cell death: the calcium-apoptosis link. Nat Rev Mol Cell Biol, 2003, 4: 552–565
Simao AM, Yadav MC, Ciancaglini P, Millan JL. Proteoliposomes as matrix vesicles’ biomimetics to study the initiation of skeletal mineralization. Braz J Med Biol Res, 2010, 43: 234–241
Simao AMS, Bolean M, Hoylaerts MF, Millan JL, Ciancaglini P. Effects of pH on the production of phosphate and pyrophosphate by matrix vesicles’ biomimetics. Calc Tissue Int, 2013, 93: 222–232
Simao AMS, Yadav MC, Narisawa S, Bolean M, Pizauro JM, Hoylaerts MF, Ciancaglini P, Millan JL. Proteoliposomes harboring alkaline phosphatase and nucleotide pyrophosphatase as matrix vesicle biomimetics. J Biol Chem, 2010, 285: 7598–7609
Zayas C, Gonzalez D, Acevedo R, del Campo J, Lastre M, Gonzalez E, Romeu B, Cuello M, Balboa J, Cabrera O, Guilherme L, Perez O. Pilot scale production of the vaccine adjuvant proteoliposome derived cochleates (AFCo1) from Neisseria meningitidis serogroup B. BMC Immunol, 2013, 14(Suppl 1): S4
Acevedo R, Perez O, Zayas C, Perez JL, Callico A, Cedre B, Garcia L, McKee D, Mullen AB, Ferro VA. Cochleates derived from Vibrio cholerae O1 proteoliposomes: the impact of structure transformation on mucosal immunisation. PLoS One, 2012, 7: e46461
Guilherme L, Postol E, de Barros SF, Higa F, Alencar R, Lastre M, Zayas C, Puschel CR, Silva WR, Sa-Rocha LC, Sa-Rocha VM, Perez O, Kalil J. A vaccine against S. pyogenes: design and experimental immune response. Methods, 2009, 49: 316–321
Acevedo R, Callico A, del Campo J, Gonzalez E, Cedre B, Gonzalez L, Romeu B, Zayas C, Lastre M, Fernandez S, Oliva R, Garcia L, Luis Perez J, Perez O. Intranasal administration of proteoliposome-derived cochleates from Vibrio cholerae O1 induce mucosal and systemic immune responses in mice. Methods, 2009, 49: 309–315
Perez O, Lastre M, Cabrera O, del Campo J, Bracho G, Cuello M, Balboa J, Acevedo R, Zayas C, Gil D, Mora N, Gonzalez D, Perez R, Gonzalez E, Barbera R, Fajardo EM, Sierra G, Solis RL, Campa C. New vaccines require potent adjuvants like AFPL1 and AFCo1. Scand J Immunol, 2007, 66: 271–277
Sobel A, Weber M, Changeux J-P. Large-scale purification of the acetylcholine-receptor protein in its membrane-bound and detergent-extracted forms from Torpedo marmorata electric organ. Eur J Biochem, 1977, 80: 215–224
Epstein M, Racker E. Reconstitution of carbamylcholine-dependent sodium ion flux and desensitization of the acetylcholine receptor from Torpedo californica. J Biol Chem, 1978, 253: 6660–6662
Schiebler W, Hucho F. Membranes rich in acetylcholine receptor: characterization and reconstitution to excitable membranes from exogenous lipids. Eur J Biochem, 1978, 85: 55–63
Ramos J, Jung WY, Ramos-Franco J, Mignery GA, Fill M. Single channel function of inositol 1,4,5-trisphosphate receptor type-1 and-2 isoform domain-swap chimeras. J Gen Physiol, 2003, 121: 399–411
Mignery GA, Johnston PA, Südhof TC. Mechanism of Ca2+ inhibition of inositol 1,4,5-trisphosphate (InsP3) binding to the cerebellar InsP3 receptor. J Biol Chem, 1992, 267: 7450–7455
Kameyama A, Shearman MS, Sekiguchi K, Kameyama M. Cyclic AMP-dependent protein kinase but not protein kinase C regulates the cardiac Ca2+ channel through phosphorylation of its alpha(1) subunit. J Biochem, 1996, 120: 170–176
Navarro J, Pyun HY, Essig A. Voltage-dependence of phosphoenzyme formation of reconstituted Ca2+-ATPase vesicles. Biophys J, 1985, 47: A345–345
Cheng KH, Lepock JR, Hui SW, Yeagle PL. The role of cholesterol in the activity of reconstituted Ca-ATPase vesicles containing unsaturated phosphatidylethanolamine. J Biol Chem, 1986, 261: 5081–5087
Wakabayashi S, Shigekawa M. Rapid reconstitution and characterization of highly-efficient sarcoplasmic-reticulum Ca pump. Biochim Biophys Acta, 1985, 813: 266–276
Moffett S, Brown DA, Linder ME. Lipid-dependent targeting of G proteins into rafts. J Biol Chem, 2000, 275: 2191–2198
Dalziel JE, Wong SS, Phung T, Zhang YL, Dunlop J. Expression of human BK ion channels in Sf9 cells, their purification using metal affinity chromatography, and functional reconstitution into planar lipid bilayers. J Chromatogr B Anal Technol Biomed Life Sci, 2007, 857: 315–321
Weber T, Zemelman BV, McNew JA, Westermann B, Gmachl M, Parlati F, Söllner TH, Rothman JE. SNAREpins: minimal machinery for membrane fusion. Cell, 1998, 92: 759–772
Nickel W, Weber T, McNew JA, Parlati F, Sollner TH, Rothman JE. Content mixing and membrane integrity during membrane fusion driven by pairing of isolated v-SNAREs and t-SNAREs. Proc Natl Acad Sci USA, 1999, 96: 12571–12576
Liu TT, Tucker WC, Bhalla A, Chapman ER, Weisshaar JC. SNARE-driven, 25-millisecond vesicle fusion in vitro. Biophys J, 2005, 89: 2458–2472
Cho WJ, Shin L, Ren G, Jena BP. Structure of membrane-associated neuronal SNARE complex: implication in neurotransmitter release. J Cell Mol Med, 2009, 13: 4161–4165
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Wang, L., Tonggu, L. Membrane protein reconstitution for functional and structural studies. Sci. China Life Sci. 58, 66–74 (2015). https://doi.org/10.1007/s11427-014-4769-0
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DOI: https://doi.org/10.1007/s11427-014-4769-0