Sat, 26 Jul 2008 17:17:06 BST CiteULike: jyuh Bernhard http://www.citeulike.org/user/jyuh/author/Bernhard 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 http://www.citeulike.org/user/jyuh/article/2463140 <i>Curr Opin Chem Biol (21 February 2008)</i><br /><br />Availability of a suite of biomarkers for early detection, stratification into distinct subtypes, and monitoring progression or response to therapy promises significant improvements in clinical outcomes for cancer patients. However, despite the recent progress in proteomics technologies based on mass spectrometry (MS), discovery of novel clinical assessment tools has been slow. This is, partly due to the inherent difficulties in working with blood as the biospecimen for candidate discovery. A better understanding of the limitations of blood for comparative protein profiling and a better appreciation of the advantages of cancer tissue or cancer cell secretomes have the potential to greatly enhance the progress. Proteomics-driven cancer biomarker discovery: looking to the future. Richard J Simpson Oliver K Bernhard David W Greening Robert L Moritz doi:10.1016/j.cbpa.2008.02.010 Curr Opin Chem Biol (21 February 2008) 2008-03-04T04:34:27-00:00 2008 Curr Opin Chem Biol 1367-5931 no-tag http://www.citeulike.org/user/jyuh/article/2170396 <i>Analytical and Bioanalytical Chemistry, Vol. 389, No. 4. (October 2007), pp. 991-1002.</i> Sample preparation by in-gel digestion for mass spectrometry-based proteomics Granvogl Bernhard Ploscher Matthias Eichacker Lutz doi:10.1007/s00216-007-1451-4 Analytical and Bioanalytical Chemistry, Vol. 389, No. 4. (October 2007), pp. 991-1002. 2007-12-26T15:41:08-00:00 2007 Analytical and Bioanalytical Chemistry 1618-2642 389 4 991 1002 Springer 2dge 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