Sat, 05 Jul 2008 00:17:40 BST CiteULike: jyuh Dötsch http://www.citeulike.org/user/jyuh/author/Dötsch CiteULike.org en-gb Copyright © 2004-2008 citeulike.org http://www.citeulike.org/user/jyuh/article/887292 <i>Methods (26 August 2006)</i><br /><br />Cell-free expression techniques have emerged as promising tools for the production of membrane proteins for structural and functional analysis. Elimination of toxic effects and a variety of options to stabilize the synthesized proteins enable the synthesis of otherwise difficult to obtain proteins. Modifications in the reaction design result in preparative scale production rates of cell-free reactions and yield in milligram amounts of membrane proteins per one millilitre of reaction volume. A diverse selection of detergents can be supplied into the reaction system without inhibitory effects to the translation machinery. This offers the unique opportunity to produce a membrane protein directly into micelles of a detergent of choice. We present detailed protocols for the cell-free production of membrane proteins in different modes and we summarize the current knowledge of this technique. A special emphasize will be on the production of soluble and functionally folded membrane proteins in presence of suitable detergents. In addition, we will highlight the advantages of cell-free expression for the structural analysis of membrane proteins especially by liquid state nuclear magnetic resonance spectroscopy and we will discuss new strategies for structural approaches. Preparative scale cell-free expression systems: New tools for the large scale preparation of integral membrane proteins for functional and structural studies. Daniel Schwarz Christian Klammt Alexander Koglin Frank Löhr Birgit Schneider Volker Dötsch Frank Bernhard doi:10.1016/j.ymeth.2006.07.001 Methods (26 August 2006) 2006-10-06T13:02:38-00:00 2006 Methods 1046-2023 no-tag http://www.citeulike.org/user/jyuh/article/2721953 <i>Cellular and molecular life sciences : CMLS (14 April 2008)</i><br /><br />The preparation of sufficient amounts of high-quality samples is still the major bottleneck for the characterization of membrane proteins by in vitro approaches. The hydrophobic nature, the requirement for complicated transport and modification pathways, and the often observed negative effects on membrane properties are intrinsic features of membrane proteins that frequently cause significant problems in overexpression studies. Establishing efficient protocols for the production of functionally folded membrane proteins is therefore a challenging task, and numerous specific characteristics have to be considered. In addition, a variety of expression systems have been developed, and choice of appropriate techniques could strongly depend on the desired target membrane proteins as well as on their intended applications. The production of membrane proteins is a highly dynamic field and new or modified approaches are frequently emerging. The review will give an overview of currently established processes for the production of functionally folded membrane proteins. Large-scale production of functional membrane proteins. F Junge B Schneider S Reckel D Schwarz V Dötsch F Bernhard doi:10.1007/s00018-008-8067-5 Cellular and molecular life sciences : CMLS (14 April 2008) 2008-04-26T15:09:16-00:00 2008 Cellular and molecular life sciences : CMLS 1420-682X no-tag