Abstract
In order to determine the structure of a protein by X-ray crystallography, well ordered three-dimensional crystals are required. However, despite the wealth of experience accumulated in the course of the crystallization and structural analyses of several hundred soluble globular proteins and their complexes, the process of crystallization still remains something of an art, and is often the rate limiting step of any analysis. For membrane proteins that present an additional challenge by virtue of their amphipathic nature, experience is considerably more limited, and the first three-dimensional crystals suitable for X-ray analysis were only reported in 1980 (1,2). Many membrane proteins form two-dimensional arrays in situ, and these may be studied by electron microscopy and electron diffraction of tilted specimens to determine their three-dimensional structure, but only in the pioneering study of bacteriorhodopsin has the resolution of the structural analysis approached that obtainable by X-ray crystallography (3). The formation of two-dimensional crystalline arrays will not be considered in this chapter. The first membrane protein crystal structure, the bacterial photosynthetic reaction center complex, was solved in 1985 (4); this was followed by the second reaction center structure in 1986 (5,6), and more recently porin in 1991 (7). Since these pioneering studies, increasing numbers of membrane protein crystallizations have been reported. For recent reviews, see refs. 8–11.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Michel, H. and Oesterhelt, D. (1980) Three-dimensional crystals of membrane proteins: Bacteriorhodopsin. Proc Natl. Acad. Sci USA 77, 1283–1285.
Garavito, R. M. and Rosenbusch, J P. (1980) Three-dimensional crystals of an integral membrane protein. J Cell Biol. 86, 327–329.
Henderson, R., Baldwin, J. M., Ceska, T. A., Zemlin, F., Beckmann, E, and Downing, K. H. (1990) Model for the structure of bacteriorhodopsin based upon high-resolution electron cryo-microscopy. J Mol Biol 213, 899–929
Deisenhofer, J., Epp, O., Miki, K., Huber, R., and Michel, H. (1985) Structure of the protein subunits in the photosynthetic reaction centre of Rhodopseudo-monas viridis at 3 Å resolution. Nature 318, 618–624
Chang, C.-H, Tiede, D., Tang, J., Smith, U., Norris, J., and Schiffer, M. (1986) Structure of Rhodopseudomonas sphaeroides R-26 reaction centre. FEBS Lett 205, 82–86.
Allen, J. P., Feher, G., Yeates, T. O., Rees, D. C, Deisenhoffer, J., Michel, H., and Huber, R. (1986) Structural homology of reaction centres from Rhodopseudomonas sphaeroides and Rhodopseudomonas viridis as determined by X-ray diffraction. Proc. Natl Acad. Sci. USA 83, 8589–8593.
Weiss, M. S., Kreusch, A., Schiltz, E., Nestel, U., Welte, W., Weckesser, J., and Schulz, G E. (1991) The structure of porin from Rhodobacter capsulatus at 1.8 Å resolution. FEBS Lett 280, 379–382.
Michel, H. (1983) Crystallisation of membrane proteins. Trends Biochem. Sci. 8, 56–59.
Garavito, R. M., Marcovic-Hously, Z., and Jenkins, J. (1986) The growth and characterisation of membrane protein crystals. J. Crystal Growth 76, 701–709.
Kühlbrandt, W. (1988) Three-dimensional crystallisation of membrane proteins. Q. Rev. Biophys 21, 429–477.
Michel, H. (1991) Crystallisation of Membrane Proteins CRC, Boca Raton, FL
Pauptit, R. A., Schirmer, T., Jansonius, J. N., Rosenbusch, J. P., Parker, M. W., Tucker, A. D, Tsernoglou, D., Weiss, M S., and Schulz, G. E. (1991) A common channel-forming motif in evolutionarily distant porins. J. Struct. Biol. 107, 136–145.
Timmins, P. A, Hauk, J., Wacker, T., and Welte, W. (1991) The influence of heptane-1,2,3-triol on the size and shape of LDAO micelles. Implications for the crystallisation of membrane proteins. FEBS Lett. 280, 115–120.
McPherson, A. (1990) Current approaches to macromolecular crystallisation. Eur J Biochem. 189, 1–23
McPherson, A. (1982) Preparation and Analysis of Protein Crystals. Wiley, New York.
Wyckoff, H. W., Hirs, C. H. W., and Timasheff, S. N. (eds.) (1985) Section II: Crystallisation and treatment of crystals, in Methods in Enzymology, vol. 114, Part A, Academic, London
Michel, H (1982) Three-dimensional crystals of a membrane protein complex. The photosynthetic reaction centre from Rhodopseudomonas viridis. J. Mol. Biol. 158, 567–572
Chang, C-H., Schiffer, M., Tiede, D., Smith, U., and Norris, J (1985) Characterisation of bacterial photosynthetic reaction centre crystals from Rhodopseudomonas sphaeroides R-26 by X-ray diffraction. J. Mol. Biol 186, 201–203.
Frank, H A, Taremi, S S.,and Knox, J. R. (1987) Crystallisation and preliminary X-ray and optical spectroscopic characterisation of the photochemical reaction centre from Rhodobacter sphaeroides strain 2 4.1. J. Mol. Biol. 198, 139–141
Welte, W., Wacker, T., Leis, M., Kreutz, W., Shiozawa, J., Gad’on, N., and Drews, G. (1985) Crystallisation of the photosynthetic light-harvesting pigment-protein complex B800-850 of Rhodopseudomonas capsulata. FEBS Lett. 182, 260–264.
Kuhlbrandt, W. (1987) Three-dimensional crystals of the light-harvesting chlorophyll a/b protein complex from pea chloroplasts. J. Mol. Biol. 194, 757–762.
Garavito, R. M, Jenkins, J, Jansonius, J N, Karlsson, R, and Rosenbusch, J. P. (1983) X-ray diffraction analysis of matrix porin, an integral membrane protein from Escherichia coli outer membranes J. Mol. Biol 164, 313–327.
Kreusch, A, Weiss, M. S., Welte, W, Weckesser, J., and Schulz, G. E. (1991) Crystals of an integral membrane protein diffracting to 1.8 A resolution. J. Mol Biol 217, 9–10.
Garavito, R. M., Hinz, U., and Neuhaus, J.-M. (1984) The crystallisation of outer membrane proteins from Escherichia coli. Studies on lamB and ompA gene products. J. Biol. Chem. 259, 4254–4257.
Stauffer, K. A., Page, M G. P., Hardmeyer, A., Keller, T A, and Pauptit, R A. (1990) Crystallisation and preliminary X-ray characterisation of maltoporin from Escherichia coli J Mol. Biol. 211, 297–299.
Tucker, A. D., Jackman, S., Parker, M. W., and Tsernoglou, D (1991) Crystallisation and preliminary X-ray analysis of phosphoporin from the outer membrane of Escherichia coli. J. Mol. Biol. 222, 881–884.
Pauptit, R. A., Zhang, H., Rummel, G., Schirmer, T., Jansonius, J N., and Rosenbusch, J. P. (1991) Trigonal crystals of porin from Escherichia coli. J. Mol Biol. 218, 505–507.
Gros, P., Groendijk, H, Drenth, J., and Hoi, W. G J. (1988) Experiments in membrane protein crystallisation. J. Crystal Growth 90, 193–200.
Yoshikawa, S., Tera, T., Takahashi, Y., Tsukihara, T, and Caughey, W. S. (1988) Crystalline cytochrome c oxidase of bovine heart mitochondrial membrane: Composition and X-ray diffraction studies. Proc. Natl Acad Sci. USA 85, 1354–1358.
Stokes, D. L, and Green, N. M. (1990) Three-dimensional crystals of CaATPase from sarcoplasmic reticulum: Symmetry and molecular packing. Biophys J. 57, 1–14
Author information
Authors and Affiliations
Rights and permissions
Copyright information
© 1994 Humana Press Inc. Totowa, NJ
About this protocol
Cite this protocol
Sutton, B.J., Sohi, M.K. (1994). Crystallization of Membrane Proteins for X-Ray Analysis. In: Biomembrane Protocols. Methods in Molecular Biology, vol 27. Springer, Totowa, NJ. https://doi.org/10.1385/0-89603-250-7:1
Download citation
DOI: https://doi.org/10.1385/0-89603-250-7:1
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-250-7
Online ISBN: 978-1-59259-514-3
eBook Packages: Springer Protocols