Sun, 27 Jul 2008 11:11:00 BST CiteULike: jyuh Koglin http://www.citeulike.org/user/jyuh/author/Koglin 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/1447701 <i>Methods, Vol. 41, No. 4. (April 2007), pp. 355-369.</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 Lohr Birgit Schneider Volker Dotsch Frank Bernhard Methods, Vol. 41, No. 4. (April 2007), pp. 355-369. 2007-07-11T04:41:17-00:00 2007 Methods 41 4 355 369 no-tag