Katzen, F., Fletcher, J.E., Yang, J.P., Kang, D., Peterson, T.C., Cappuccio, J.A., Blanchette, C.D., Sulchek, T., Chromy,
B.A., Hoeprich, P.D., Coleman, M.A., Kudlicki, W. (2008) Insertion of membrane proteins into discoidal membranes
using a cell-free protein expression approach. J. Proteome Res. 7:3535–3542.
Keller, T., Schwarz, D., Bernhard, F., Dötsch, V., Hunte, C., Gorboulev, V., Koepsell, H. (2008) Cell-free expression and
functional reconstitution of eukaryotic drug transporters. Biochemistry 47:4552–4564.
Kigawa, T. (2010) Cell-free protein production system with the E. coli crude extract for determination of protein folds.
Methods Mol. Biol. 607:101–111.
Kigawa, T., Yabuki, T., Yoshida, Y., Tsutsui, M., Ito, Y., Shibata, T., Yokoyama, S. (1999) Cell-free production and
stable-isotope labeling of milligram quantities of proteins. FEBS Lett. 442:15–19.
Kim, T.W., Keum, J.W., Oh, I.S., Choi, C.Y., Park, C.G., Kim, D.M. (2006) Simple procedures for the construction of a
robust and cost-effective cell-free protein synthesis system. J. Biotechnol. 126:554–561.
Klammt, C., Lohr, F., Schäfer, B., Haase, W., Dötsch, V., Ruterjans, H., Glaubitz, C., Bernhard, F. (2004) High level
cell-free expression and specific labeling of integral membrane proteins. Eur. J. Biochem. 271:568–580.
Klammt, C., Schwarz, D., Fendler, K., Haase, W., Dötsch, V., Bernhard, F. (2005) Evaluation of detergents for the
soluble expression of α-helical and β-barrel-type integral membrane proteins by a preparative scale individual cellfree expression system. FEBS J. 272:6024–6038
Klammt, C., Schwarz, D., Löhr, F., Schneider, B., Dötsch, V., Bernhard, F. (2006) Cell-free expression as an emerging
technique for the large scale production of integral membrane protein. FEBS J. 273:4141–4153.
Klammt, C., Perrin, M.-H., Maslennikov, I., Renault, L., Krupa, M., Kwiatkowski, W., Stahlberg, H., Vale, W., Choe,
S. (2011) Polymer-based cell-free expression of ligand-binding family B G protein-coupled receptors without
detergents. Prot. Sci. 20:1030–1041.
Kleinschmidt, J.H. (2015) Folding of β-barrel membrane proteins in lipid bilayers – Unassisted and assisted folding and
insertion. Biochim. Biophys. Acta 1848:1927–1943.
Kleinschmidt, J.H., Popot, J.-L. (2014) Folding and stability of integral membrane proteins in amphipols. Arch. Biochem.
Biophys. 564:327–343.
Kuruma, Y., Ueda, T. (2015) The PURE system for the cell-free synthesis of membrane proteins. Nat. Protoc.
10:1328–1344.
Kuruma, Y., Suzuki, T., Ueda, T. (2010) Production of multi-subunit complexes on liposome through an E. coli cell-free
expression system. Methods Mol. Biol. 607:161–171.
LaGuerre, A., Löhr, F., Bernhard, F., Dötsch, V. (2015) Labeling of membrane proteins by cell-free expression. Meth.
Enzymol. 563:367–388.
Lyukmanova, E.N., Shenkarev, Z.O., Khabibullina, N.F., Kopeina, G.S., Shulepko, M.A., Paramonov, A.S., Mineev, K.
S., Tikhonov, R.V., Shingarova, L.N., Petrovskaya, L.E., Dolgikh, D.A., Arseniev, A.S., Kirpichnikov, M.P. (2012)
Lipid-protein nanodiscs for cell-free production of integral membrane proteins in a soluble and folded state:
Comparison with detergent micelles, bicelles and liposomes. Biochim. Biophys. Acta 1818:349–358.
Maeda, S., Schertler, G.F.X. (2013) Production of GPCR and GPCR complexes for structure determination. Curr. Opin.
Struct. Biol. 23:381–392.
Maslennikov, I., Klammt, C., Hwang, E., Kefala, G., Okamura, M., Esquivies, L., Mörs, K., Glaubitz, C., Kwiatkowski,
W., Jeon, Y.H., Choe, S. (2010) Membrane domain structures of three classes of histidine kinase receptors by cellfree expression and rapid NMR analysis. Proc. Natl. Acad. Sci. USA 107:10902–10907.
Matsubayashi, H., Kuruma, Y., Ueda, T. (2014) In vitro synthesis of the E. coli Sec translocon from DNA. Angew. Chem.
Int. Ed. Engl. 53:7535–7538.
Nirenberg, M.W., Matthaei, J.H. (1961) The dependence of cell-free protein synthesis in E. coli upon naturally occurring
or synthetic polyribonucleotides. Proc. Natl. Acad. Sci. USA 47:1588–1602.
Niwa, T., Sasaki, Y., Uemura, E., Nakamura, S., Akiyama, M., Ando, M., Sawada, S., Mukai, S.A., Ueda, T., Taguchi,
H., Akiyoshi, K. (2015) Comprehensive study of liposome-assisted synthesis of membrane proteins using a
reconstituted cell-free translation system. Sci. Rep. 5:18025.
Otzen, D.E., Andersen, K.K. (2013) Folding of outer membrane proteins. Arch. Biochem. Biophys. 531:34–43.
Park, K.-H., Berrier, C., Lebaupain, F., Pucci, B., Popot, J.-L., Ghazi, A., Zito, F. (2007) Fluorinated and hemifluorinated
surfactants as alternatives to detergents for membrane protein cell-free synthesis. Biochem. J. 403:183–187.
Park, K.-H., Billon-Denis, E., Dahmane, T., Lebaupain, F., Pucci, B., Breyton, C., Zito, F. (2011) In the cauldron of cellfree synthesis of membrane proteins: Playing with new surfactants. New Biotech. 28:255–261.
Periasamy, A., Shadiac, N., Amalraj, A., Garajová, S., Nagarajan, Y., Waters, S., Mertens, H.D.T., Hrmova, M. (2013)
Cell-free protein synthesis of membrane (1,3)-β-D-glucan (curdlan) synthase: co-translational insertion in liposomes
and reconstitution in nanodiscs. Biochim. Biophys. Acta 1828:743–757.
Picard, M., Dahmane, T., Garrigos, M., Gauron, C., Giusti, F., le Maire, M., Popot, J.-L., Champeil, P. (2006) Protective
and inhibitory effects of various types of amphipols on the Ca
2+ -ATPase from sarcoplasmic reticulum: a comparative
study. Biochemistry 45:1861–1869.
Popot, J.L. (2014) Folding membrane proteins in vitro: A table and some comments. Arch. Biochem. Biophys.
564:314–326.
378
7 Amphipol-Assisted Cell-Free Expression of Membrane Proteins
B.A., Hoeprich, P.D., Coleman, M.A., Kudlicki, W. (2008) Insertion of membrane proteins into discoidal membranes
using a cell-free protein expression approach. J. Proteome Res. 7:3535–3542.
Keller, T., Schwarz, D., Bernhard, F., Dötsch, V., Hunte, C., Gorboulev, V., Koepsell, H. (2008) Cell-free expression and
functional reconstitution of eukaryotic drug transporters. Biochemistry 47:4552–4564.
Kigawa, T. (2010) Cell-free protein production system with the E. coli crude extract for determination of protein folds.
Methods Mol. Biol. 607:101–111.
Kigawa, T., Yabuki, T., Yoshida, Y., Tsutsui, M., Ito, Y., Shibata, T., Yokoyama, S. (1999) Cell-free production and
stable-isotope labeling of milligram quantities of proteins. FEBS Lett. 442:15–19.
Kim, T.W., Keum, J.W., Oh, I.S., Choi, C.Y., Park, C.G., Kim, D.M. (2006) Simple procedures for the construction of a
robust and cost-effective cell-free protein synthesis system. J. Biotechnol. 126:554–561.
Klammt, C., Lohr, F., Schäfer, B., Haase, W., Dötsch, V., Ruterjans, H., Glaubitz, C., Bernhard, F. (2004) High level
cell-free expression and specific labeling of integral membrane proteins. Eur. J. Biochem. 271:568–580.
Klammt, C., Schwarz, D., Fendler, K., Haase, W., Dötsch, V., Bernhard, F. (2005) Evaluation of detergents for the
soluble expression of α-helical and β-barrel-type integral membrane proteins by a preparative scale individual cellfree expression system. FEBS J. 272:6024–6038
Klammt, C., Schwarz, D., Löhr, F., Schneider, B., Dötsch, V., Bernhard, F. (2006) Cell-free expression as an emerging
technique for the large scale production of integral membrane protein. FEBS J. 273:4141–4153.
Klammt, C., Perrin, M.-H., Maslennikov, I., Renault, L., Krupa, M., Kwiatkowski, W., Stahlberg, H., Vale, W., Choe,
S. (2011) Polymer-based cell-free expression of ligand-binding family B G protein-coupled receptors without
detergents. Prot. Sci. 20:1030–1041.
Kleinschmidt, J.H. (2015) Folding of β-barrel membrane proteins in lipid bilayers – Unassisted and assisted folding and
insertion. Biochim. Biophys. Acta 1848:1927–1943.
Kleinschmidt, J.H., Popot, J.-L. (2014) Folding and stability of integral membrane proteins in amphipols. Arch. Biochem.
Biophys. 564:327–343.
Kuruma, Y., Ueda, T. (2015) The PURE system for the cell-free synthesis of membrane proteins. Nat. Protoc.
10:1328–1344.
Kuruma, Y., Suzuki, T., Ueda, T. (2010) Production of multi-subunit complexes on liposome through an E. coli cell-free
expression system. Methods Mol. Biol. 607:161–171.
LaGuerre, A., Löhr, F., Bernhard, F., Dötsch, V. (2015) Labeling of membrane proteins by cell-free expression. Meth.
Enzymol. 563:367–388.
Lyukmanova, E.N., Shenkarev, Z.O., Khabibullina, N.F., Kopeina, G.S., Shulepko, M.A., Paramonov, A.S., Mineev, K.
S., Tikhonov, R.V., Shingarova, L.N., Petrovskaya, L.E., Dolgikh, D.A., Arseniev, A.S., Kirpichnikov, M.P. (2012)
Lipid-protein nanodiscs for cell-free production of integral membrane proteins in a soluble and folded state:
Comparison with detergent micelles, bicelles and liposomes. Biochim. Biophys. Acta 1818:349–358.
Maeda, S., Schertler, G.F.X. (2013) Production of GPCR and GPCR complexes for structure determination. Curr. Opin.
Struct. Biol. 23:381–392.
Maslennikov, I., Klammt, C., Hwang, E., Kefala, G., Okamura, M., Esquivies, L., Mörs, K., Glaubitz, C., Kwiatkowski,
W., Jeon, Y.H., Choe, S. (2010) Membrane domain structures of three classes of histidine kinase receptors by cellfree expression and rapid NMR analysis. Proc. Natl. Acad. Sci. USA 107:10902–10907.
Matsubayashi, H., Kuruma, Y., Ueda, T. (2014) In vitro synthesis of the E. coli Sec translocon from DNA. Angew. Chem.
Int. Ed. Engl. 53:7535–7538.
Nirenberg, M.W., Matthaei, J.H. (1961) The dependence of cell-free protein synthesis in E. coli upon naturally occurring
or synthetic polyribonucleotides. Proc. Natl. Acad. Sci. USA 47:1588–1602.
Niwa, T., Sasaki, Y., Uemura, E., Nakamura, S., Akiyama, M., Ando, M., Sawada, S., Mukai, S.A., Ueda, T., Taguchi,
H., Akiyoshi, K. (2015) Comprehensive study of liposome-assisted synthesis of membrane proteins using a
reconstituted cell-free translation system. Sci. Rep. 5:18025.
Otzen, D.E., Andersen, K.K. (2013) Folding of outer membrane proteins. Arch. Biochem. Biophys. 531:34–43.
Park, K.-H., Berrier, C., Lebaupain, F., Pucci, B., Popot, J.-L., Ghazi, A., Zito, F. (2007) Fluorinated and hemifluorinated
surfactants as alternatives to detergents for membrane protein cell-free synthesis. Biochem. J. 403:183–187.
Park, K.-H., Billon-Denis, E., Dahmane, T., Lebaupain, F., Pucci, B., Breyton, C., Zito, F. (2011) In the cauldron of cellfree synthesis of membrane proteins: Playing with new surfactants. New Biotech. 28:255–261.
Periasamy, A., Shadiac, N., Amalraj, A., Garajová, S., Nagarajan, Y., Waters, S., Mertens, H.D.T., Hrmova, M. (2013)
Cell-free protein synthesis of membrane (1,3)-β-D-glucan (curdlan) synthase: co-translational insertion in liposomes
and reconstitution in nanodiscs. Biochim. Biophys. Acta 1828:743–757.
Picard, M., Dahmane, T., Garrigos, M., Gauron, C., Giusti, F., le Maire, M., Popot, J.-L., Champeil, P. (2006) Protective
and inhibitory effects of various types of amphipols on the Ca
2+ -ATPase from sarcoplasmic reticulum: a comparative
study. Biochemistry 45:1861–1869.
Popot, J.L. (2014) Folding membrane proteins in vitro: A table and some comments. Arch. Biochem. Biophys.
564:314–326.
378
7 Amphipol-Assisted Cell-Free Expression of Membrane Proteins