TAILIEUCHUNG - Báo cáo khoa học: Functional analysis of cell-free-produced human endothelin B receptor reveals transmembrane segment 1 as an essential area for ET-1 binding and homodimer formation

The functional and structural characterization of G-protein-coupled recep-tors (GPCRs) still suffers from tremendous difficulties during sample preparation. Cell-free expression has recently emerged as a promising alter-native approach for the synthesis of polytopic integral membrane proteins and, in particular, for the production of G-protein-coupled receptors. | ỊFEBS Journal Functional analysis of cell-free-produced human endothelin B receptor reveals transmembrane segment 1 as an essential area for ET-1 binding and homodimer formation Christian Klammt1 Ankita Srivastava2 Nora Eifler3 Friederike Junge1 Michael Beyermann4 Daniel Schwarz1 Hartmut Michel2 Volker Doetsch1 and Frank Bernhard1 1 Centre for Biomolecular Magnetic Resonance Institute for BiophysicalChemistry University of Frankfurt Main Germany 2 Max-Planck-Institute for Biophysics Department of Molecular Membrane Biology Frankfurt Main Germany 3 . Mueller Institute for Microscopy Biocentre University of Basel Switzerland 4 Leibniz-Institute of Molecular Pharmacology Department of Peptide Chemistry Biochemistry Berlin Germany Keywords cell-free expression detergent micelles endothelin-1 ligand-binding site G-protein coupled receptor single-particle analysis Correspondence F. Bernhard Centre for Biomolecular Magnetic Resonance Institute for BiophysicalChemistry University of Frankfurt Main Max-von-Laue-Str. 9 D-60438 Frankfurt Main Germany Fax 49 69 798 29632 Tel 49 69 798 29620 E-mail fbern@ Received 27 March 2007 revised 26 April 2007 accepted 27 April 2007 doi The functional and structural characterization of G-protein-coupled receptors GPCRs still suffers from tremendous difficulties during sample preparation. Cell-free expression has recently emerged as a promising alternative approach for the synthesis of polytopic integral membrane proteins and in particular for the production of G-protein-coupled receptors. We have now analyzed the quality and functional folding of cell-free produced human endothelin type B receptor samples as an example of the rhodopsin-type family of G-protein-coupled receptors in correlation with different cell-free expression modes. Human endothelin B receptor was cell-free produced as a precipitate and subsequently solubilized in detergent or was directly synthesized in micelles .

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