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The 2DX robot : a membrane protein 2D crystallization Swiss Army knife

Iacovache, I. and Biasini, M. and Kowal, J. and Kukulski, W. and Chami, M. and van der Goot, F. G. and Engel, A. and Remigy, H. W.. (2009) The 2DX robot : a membrane protein 2D crystallization Swiss Army knife. Journal of Structural Biology, 169 (3). pp. 370-378.

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Official URL: http://edoc.unibas.ch/dok/A5262453

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Abstract

Among the state-of-the-art techniques that provide experimental information at atomic scale for membrane proteins, electron crystallography, atomic force microscopy and solid state NMR make use of two-dimensional crystals. We present a cyclodextrin-driven method for detergent removal implemented in a fully automated robot. The kinetics of the reconstitution processes is precisely controlled, because the detergent complexation by cyclodextrin is of stoichiometric nature. The method requires smaller volumes and lower protein concentrations than established 2D crystallization methods, making it possible to explore more conditions with the same amount of protein. The method yielded highly ordered 2D crystals diffracting to high resolution from the pore-forming toxin Aeromonas hydrophila aerolysin (2.9A), the plant aquaporin SoPIP2;1 (3.1A) and the human aquaporin-8 (hAQP8; 3.3A). This new method outperforms traditional 2D crystallization approaches in terms of accuracy, flexibility, throughput, and allows the usage of detergents having low critical micelle concentration (CMC), which stabilize the structure of membrane proteins in solution.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Former Organization Units Biozentrum > Structural Biology (Engel)
UniBasel Contributors:Engel, Andreas H and Kowal, Julia and Biasini, Marco and Chami, Mohamed
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Elsevier
ISSN:1047-8477
e-ISSN:1095-8657
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:21 Mar 2023 13:19
Deposited On:22 Mar 2012 13:30

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