Protocol prepared by Marina Casiraghi and Laurent J. Catoire on the basis of Damian et al.
(2012), Casiraghi et al. (2016), and laboratory notes.
References
Agosto, M.A., Zhang, Z., He, F., Anastassov, I.A., Wright, S.J., McGehee, J., Wensel, T.G. (2014) Oligomeric state of
purified TRPM1, a protein essential for dim light vision. J. Biol. Chem. 289:27019–27033.
Althoff, T., Mills, D.J., Popot, J.-L., Kühlbrandt, W. (2011) Assembly of electron transport chain components in bovine
mitochondrial supercomplex I 1 III 2 IV 1 . EMBO J. 30:4652–4664.
Arunmanee, W., Harris, J.R., Lakey, J.H. (2014) Outer membrane protein F stabilised with minimal amphipol forms
linear arrays and LPS-dependent 2D crystals. J. Membr. Biol. 247:949–956.
Baenziger, J.E., Hénault, C.M., Therien, J.P.D., Sun, J. (2015) Nicotinic acetylcholine receptor-lipid interactions:
Mechanistic insight and biological function. Biochim. Biophys. Acta 1848:1806–1817.
Bai, X.-C., Yan, C., Yang, G., Lu, P., Ma, D., Sun, L., Zhou, R., Scheres, S.H.W., Shi, Y. (2015) An atomic structure of
human γ-secretase. Nature 525:212–218.
Baker, M.R., Fan, G., Serysheva, I.I. (2015) Single-particle cryo-EM of the ryanodine receptor channel in an aqueous
environment. Eur. J. Transl. Myol. 25:35–48.
Banères, J.-L., Popot, J.-L., Mouillac, B. (2011) New advances in production and functional folding of G protein-coupled
receptors. Trends Biotechnol. 29:314–322.
Basit, H., Sharma, S., Van der Heyden, A., Gondran, C., Breyton, C., Dumy, P., Winnik, F.M., Labbé, P. (2012)
Amphipol-mediated surface immobilization of FhuA: a platform for label-free detection of the bacteriophage protein
pb5. Chem. Commun. 48:6037–6039.
Bazzacco, P. (2009) Non-ionic amphipols: new tools for in vitro studies of membrane proteins. Validation and
development of biochemical and biophysical applications. Thèse de Doctorat, Université Paris-7, Paris, 176 p.
Fig. 5.44 Characterization of MSP1D1 NDs hosting the leukotriene BLT2 receptor. (A) Electron
microscopy image of negatively stained BLT2/ND complexes purified by SEC (cf. Fig. 5.43). The discs
are 12–15 nm in diameter. White arrows, top views; black arrows, side views. (B) SDS-polyacrylamide gel
electrophoresis analysis. BLT2/NDs, complexes purified by SEC; BLT2/SDS and MSP1D1, controls.
Note that, as frequently occurs with polytopic MPs, BLT2 tends to form oligomers in SDS (Reprinted with
permission from Casiraghi et al. 2016, # 2016 American Chemical Society).
322
5 Formation and Properties of Membrane Protein/Amphipol Complexes
(2012), Casiraghi et al. (2016), and laboratory notes.
References
Agosto, M.A., Zhang, Z., He, F., Anastassov, I.A., Wright, S.J., McGehee, J., Wensel, T.G. (2014) Oligomeric state of
purified TRPM1, a protein essential for dim light vision. J. Biol. Chem. 289:27019–27033.
Althoff, T., Mills, D.J., Popot, J.-L., Kühlbrandt, W. (2011) Assembly of electron transport chain components in bovine
mitochondrial supercomplex I 1 III 2 IV 1 . EMBO J. 30:4652–4664.
Arunmanee, W., Harris, J.R., Lakey, J.H. (2014) Outer membrane protein F stabilised with minimal amphipol forms
linear arrays and LPS-dependent 2D crystals. J. Membr. Biol. 247:949–956.
Baenziger, J.E., Hénault, C.M., Therien, J.P.D., Sun, J. (2015) Nicotinic acetylcholine receptor-lipid interactions:
Mechanistic insight and biological function. Biochim. Biophys. Acta 1848:1806–1817.
Bai, X.-C., Yan, C., Yang, G., Lu, P., Ma, D., Sun, L., Zhou, R., Scheres, S.H.W., Shi, Y. (2015) An atomic structure of
human γ-secretase. Nature 525:212–218.
Baker, M.R., Fan, G., Serysheva, I.I. (2015) Single-particle cryo-EM of the ryanodine receptor channel in an aqueous
environment. Eur. J. Transl. Myol. 25:35–48.
Banères, J.-L., Popot, J.-L., Mouillac, B. (2011) New advances in production and functional folding of G protein-coupled
receptors. Trends Biotechnol. 29:314–322.
Basit, H., Sharma, S., Van der Heyden, A., Gondran, C., Breyton, C., Dumy, P., Winnik, F.M., Labbé, P. (2012)
Amphipol-mediated surface immobilization of FhuA: a platform for label-free detection of the bacteriophage protein
pb5. Chem. Commun. 48:6037–6039.
Bazzacco, P. (2009) Non-ionic amphipols: new tools for in vitro studies of membrane proteins. Validation and
development of biochemical and biophysical applications. Thèse de Doctorat, Université Paris-7, Paris, 176 p.
Fig. 5.44 Characterization of MSP1D1 NDs hosting the leukotriene BLT2 receptor. (A) Electron
microscopy image of negatively stained BLT2/ND complexes purified by SEC (cf. Fig. 5.43). The discs
are 12–15 nm in diameter. White arrows, top views; black arrows, side views. (B) SDS-polyacrylamide gel
electrophoresis analysis. BLT2/NDs, complexes purified by SEC; BLT2/SDS and MSP1D1, controls.
Note that, as frequently occurs with polytopic MPs, BLT2 tends to form oligomers in SDS (Reprinted with
permission from Casiraghi et al. 2016, # 2016 American Chemical Society).
322
5 Formation and Properties of Membrane Protein/Amphipol Complexes