Abstract
This method of two-dimensional crystallization of proteins on a lipid monolayer aims at producing 2D crystals of membrane proteins, which can provide structural information at high resolution by electron crystallography.
A lipid monolayer is spread over the whole air–water interface of a drop, which provides a substrate for protein binding. The protein of interest is then adsorbed onto the lipid monolayer and forms a closely packed layer. The reconstitution step of the membrane protein into a lipid bilayer is realized by elimination of detergent. The combined effect of the elevated protein concentration, the alignment of the protein on the lipid monolayer, and the fluid monolayer film properties are conducive, in some cases, to the formation of 2D crystals. The use of locally fluorinated lipids in avoiding solubilization of the lipid monolayer by detergents is presented and discussed.
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References
Fromherz P (1971) Electron microsopic studies of lipid protein films. Nature 231:267–268
Lebeau L, Schultz P, Célia H, Mésini P, Nuss S, Klinger C, Olland S, Oudet P, Mioskowski C (1996) Specifically designed lipid assemblies as tools for two-dimensional crystallization of soluble biological macromolecules. In: Barenholz Y, Lasic DD (eds) Handbook of non-medical applications of liposomes. Vol. II. CRC, Boca Raton, pp 153–186
Dietrich J, Venien-Bryan C (2005) Strategies for two-dimensional crystallization of proteins using lipid monolayers. Imperial College Press, London
Nuss S, Mioskowski C, Lebeau L (1999) Synthesis of new fluidity-enhanced amphiphilic compounds for soluble protein two-dimensional crystallization purpose. Chem Phys Lipids 103:21–35
Lebeau L, Nuss S, Schultz P, Oudet P, Mioskowski C (1999) Self-assembly of soluble proteins on functionalized lipid layers: a tentative correlation between the fluidity properties of the lipid film and protein ordering. Chem Phys Lipids 103:37–46
Courty S, Lebeau L, Martel L, Lenne PF, Balavoine F, Dischert W, Konovalov O, Mioskowski C, Legrand JF, Venien-Bryan C (2002) Two-dimensional crystallization of a His-tagged protein on monolayers of fluidity-enhanced Ni2+-chelating lipids. Langmuir 18:9502–9512
Fontaine P, Faure MC, Muller F, Poujade M, Micha JS, Rieutord F, Goldmann M (2007) Unexpected stability of phospholipid langmuir monolayers deposited on triton X-100 aqueous solutions. Langmuir 23:12959–12965
Levy D, Mosser G, Lambert O, Moeck GS, Bald D, Rigaud JL (1999) Two-dimensional crystallization on lipid layer: a successful approach for membrane proteins. J Struct Biol 127:44–52
Lata S, Reichel A, Brock R, Tampé R, Piehler J (2005) High-affinity adaptors for switchable recognition of histidine-tagged proteins. J Am Chem Soc 127:10205–10215
Lebeau L, Lach F, Venien-Bryan C, Renault A, Dietrich J, Jahn T, Palmgren MG, Kuhlbrandt W, Mioskowski C (2001) Two-dimensional crystallization of a membrane protein on a detergent-resistant lipid monolayer. J Mol Biol 308:639–647
Dauvergne J, Polidori A, Vénien-Bryan C, Pucci B (2008) Synthesis of a hemifluorinated amphiphile designed for self-assembly and two-dimensional crystallization of membrane protein. Tetrahedron Lett 49:2247–2250
Hussein WM, Ross BP, Landsberg MJ, Levy D, Hankamer B, McGeary RP (2009) Synthesis of nickel-chelating fluorinated lipids for protein monolayer crystallizations. J Org Chem 74:1473–1479
Hussein WM, Ross BP, Landsberg MJ, Hankamer B, McGeary RP (2009) Synthetic approaches to functionalized lipids for protein monolayer crystallizations. Curr Org Chem 13:1378–1405
Vénien-Bryan C, Lenne PF, Zakri C, Renault A, Brisson A, Legrand JF, Berge B (1998) Characterization of the growth of 2D protein crystals on a lipid monolayer by ellipsometry and rigidity measurements coupled to electron microscopy. Biophys J 74:2649–2657
Lenne PF, Berge B, Renault A, Zakri C, Venien-Bryan C, Courty S, Balavoine F, Bergsma-Schutter W, Brisson A, Grubel G, Boudet N, Konovalov O, Legrand JF (2000) Synchrotron radiation diffraction from two-dimensional protein crystals at the air/water interface. Biophys J 79:496–500
Darst SA, Ahlers M, Meller PH, Kubalek EW, Blankenburg R, Ribi HO, Ringsdorf H, Kornberg RD (1991) Two-dimensional crystals of streptavidin on biotinylated lipid layers and their interactions with biotinylated macromolecules. Biophys J 59:387–396
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Lebeau, L., Vénien-Bryan, C. (2013). Monolayer Two-Dimensional Crystallization of Membrane Proteins. In: Schmidt-Krey, I., Cheng, Y. (eds) Electron Crystallography of Soluble and Membrane Proteins. Methods in Molecular Biology, vol 955. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-176-9_4
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DOI: https://doi.org/10.1007/978-1-62703-176-9_4
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