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Membrane protein crystallization

Published on Apr 1, 2003in Journal of Structural Biology3.75
· DOI :10.1016/S1047-8477(03)00043-1
Martin Caffrey56
Estimated H-index: 56
(OSU: Ohio State University)
Abstract
Abstract The need for high-resolution structure information on membrane proteins is immediate and growing. Currently, the only reliable way to get it is crystallographically. The rate-limiting step from protein to structure is crystal production. An overview of the current ideas and experimental approaches prevailing in the area of membrane protein crystallization is presented. The long-established surfactant-based method has been reviewed extensively and is not examined in detail here. The focus instead is on the latest methods, all of which exploit the spontaneous self-assembling properties of lipids and detergent as vesicles (vesicle-fusion method), discoidal micelles (bicelle method), and liquid crystals or mesophases (in meso or cubic-phase method). In the belief that a knowledge of the underlying phase science is integral to understanding the molecular basis of these assorted crystallization strategies, the article begins with a brief primer on lipids, mesophases, and phase science, and the related issue of form and function as applied to lipids is addressed. The experimental challenges associated with and the solutions for procuring adequate amounts of homogeneous membrane proteins, or parts thereof, are examined. The cubic-phase method is described from the following perspectives: how it is done in practice, its general applicability and successes to date, and the nature of the mesophases integral to the process. Practical aspects of the method are examined with regard to salt, detergent, and screen solution effects; crystallization at low temperatures; tailoring the cubic phase to suit the target protein; different cubic-phase types; dealing with low-protein samples, colorless proteins, microcrystals, and radiation damage; transport within the cubic phase for drug design, cofactor retention, and phasing; using spectroscopy for quality control; harvesting crystals; and miniaturization and robotization for high-throughput screening. The section ends with a hypothesis for nucleation and growth of membrane protein crystals in meso. Thus far, the bicelle and vesicle-fusion methods have produced crystals of one membrane protein, bacteriorhodopsin. The experimental details of both methods are reviewed and their general applicability in the future is commented on. The three new methods are rationalized by analogy to crystallization in microgravity and with respect to epitaxy. A list of Web resources in the area of membrane protein crystallogenesis is included.
  • References (84)
  • Citations (281)
References84
Newest
#1Raimond B G Ravelli R B G (EMBL-EBI: European Bioinformatics Institute)H-Index: 42
#2Hanna-Kirsti S. Leiros (University of Tromsø)H-Index: 21
Last.Sean McSweeney (European Synchrotron Radiation Facility)H-Index: 42
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#1Etana Padan (HUJI: Hebrew University of Jerusalem)H-Index: 54
#2Hunte (MPG: Max Planck Society)H-Index: 1
Last.Reilaender (MPG: Max Planck Society)H-Index: 1
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#1Carola Hunte (MPG: Max Planck Society)H-Index: 36
#2Hartmut Michel (MPG: Max Planck Society)H-Index: 70
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#2Leilei Zhang (CNRS: Centre national de la recherche scientifique)H-Index: 1
Last.Nguyen M (CNRS: Centre national de la recherche scientifique)H-Index: 14
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#1Thomas G. Meikle (RMIT: RMIT University)H-Index: 1
#2Ashish Sethi (University of Melbourne)H-Index: 4
Last.Shenggen Yao (University of Melbourne)H-Index: 25
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#1Raffaele Mezzenga (ETH Zurich)H-Index: 56
#2John M. Seddon (Imperial College London)H-Index: 36
Last.Laurent Sagalowicz (Nestlé)H-Index: 27
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#1Saki Fujiwara (TUAT: Tokyo University of Agriculture and Technology)H-Index: 3
#2Hiroyuki Ohno (TUAT: Tokyo University of Agriculture and Technology)H-Index: 65
Last.Takahiro Ichikawa (TUAT: Tokyo University of Agriculture and Technology)H-Index: 19
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