Preparative scale cell-free expression systems: New tools for the large scale preparation of integral membrane proteins for functional and structural studies.

@article{citeulike:887292, abstract = {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.}, address = {Centre for Biomolecular Magnetic Resonance, University of Frankfurt/Main, Institute for Biophysical Chemistry, Max-von-Laue-Str. 9, D-60438 Frankfurt/Main, Germany.}, author = {Schwarz, Daniel and Klammt, Christian and Koglin, Alexander and L\"{o}hr, Frank and Schneider, Birgit and D\"{o}tsch, Volker and Bernhard, Frank }, citeulike-article-id = {887292}, doi = {10.1016/j.ymeth.2006.07.001}, issn = {1046-2023}, journal = {Methods}, keywords = {cell\_free, membrane\_protein}, month = {August}, posted-at = {2008-07-24 08:46:22}, priority = {2}, title = {Preparative scale cell-free expression systems: New tools for the large scale preparation of integral membrane proteins for functional and structural studies.}, url = {http://dx.doi.org/10.1016/j.ymeth.2006.07.001}, year = {2006} }

TY - JOUR ID - citeulike:887292 L3 - citeulike-article-id:887292 TI - Preparative scale cell-free expression systems: New tools for the large scale preparation of integral membrane proteins for functional and structural studies. JF - Methods AD - Centre for Biomolecular Magnetic Resonance, University of Frankfurt/Main, Institute for Biophysical Chemistry, Max-von-Laue-Str. 9, D-60438 Frankfurt/Main, Germany. SN - 1046-2023 N2 - 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. KW - cell_free KW - membrane_protein AU - Schwarz, Daniel AU - Klammt, Christian AU - Koglin, Alexander AU - Löhr, Frank AU - Schneider, Birgit AU - Dötsch, Volker AU - Bernhard, Frank PY - 2006/08/26/ UR - http://dx.doi.org/10.1016/j.ymeth.2006.07.001 ER -