TAILIEUCHUNG - Báo cáo khoa học: Evaluation of detergents for the soluble expression of a-helical and b-barrel-type integral membrane proteins by a preparative scale individual cell-free expression system

Cell-free expression has become a highly promising tool for the fast and efficient production of integral membrane proteins. The proteins can be produced as precipitates that solubilize in mild detergents usually without any prior denaturation steps. Alternatively, membrane proteins can be syn-thesized in a soluble form by adding detergents to the cell-free system. | iFEBS Journal Evaluation of detergents for the soluble expression of a-helical and b-barrel-type integral membrane proteins by a preparative scale individual cell-free expression system Christian Klammt1 z Daniel Schwarz1 z Klaus Fendler2 Winfried Haase3 Volker Dotsch1 and Frank Bernhard1 1 Centre for Biomolecular Magnetic Resonance University of Frankfurt Main Institute for BiophysicalChemistry Frankfurt Main Germany 2 Max-Planck-Institute for Biophysics Department for BiophysicalChemistry Frankfurt Main Germany 3 Max-Planck-Institute for Biophysics Department for StructuralBiology Frankfurt Main Germany Keywords cellfree membrane protein synthesis detergent micelles functionalp-barrel membrane protein G-protein coupled receptor transporter Correspondence F. Bernhard Centre for Biomolecular Magnetic Resonance University of Frankfurt Main Institute for Biophysical Chemistry Marie-Curie-Str. 9 D-60439 Frankfurt Main Germany Fax 49 69 798 29632 Tel 49 69 798 29620 E-mail fbern@ These authors contributed equally to this manuscript. Received 1 July 2005 revised 26 September 2005 accepted 3 October 2005 doi Cell-free expression has become a highly promising tool for the fast and efficient production of integral membrane proteins. The proteins can be produced as precipitates that solubilize in mild detergents usually without any prior denaturation steps. Alternatively membrane proteins can be synthesized in a soluble form by adding detergents to the cell-free system. However the effects of a representative variety of detergents on the production solubility and activity of a wider range of membrane proteins upon cell-free expression are currently unknown. We therefore analyzed the cell-free expression of three structurally very different membrane proteins namely the bacterial a-helical multidrug transporter EmrE the p-barrel nucleoside transporter Tsx and the porcine vasopressin receptor of the eukaryotic superfamily of

• Báo cáo khoa học
• Report medicine
• traditional medicine
• scientific reports
• biochemical studies
• environmental medicine
• Crystal structure
• medical knowledge
• medical research
• analysis of cytoplasmic
• oxidation
• bacteria
• hemoglobin
• medicine
• cell proliferation
• breast cancer
• chemical reaction
• gastric cancer
• hormone medicine
• tumor cells