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Isotope labeling of mammalian GPCRs in HEK293 cells and characterization of the C-terminus of bovine rhodopsin by high resolution liquid NMR spectroscopy |
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| Authors: | Karla Werner Christian Richter Judith Klein-Seetharaman Harald Schwalbe |
| Institution: | (1) Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe-University, Max-von-Laue-Strasse 7, Frankfurt/Main, D-60438, Germany;(2) Department of Structural Biology, University of Pittsburgh School of Medicine, Biomedical Science Tower 3, Rm 2051, Fifth Avenue, Pittsburgh, PA 15261, USA |
| Abstract: | High amino acid coverage labeling of the mammalian G protein coupled receptors (GPCR) rhodopsin was established with 15N and 15N/13C isotopes. Rhodopsin was expressed at preparative scale in HEK293S cells and studied in full-length by NMR spectroscopy in detergent micelle solution. This resulted in the assignment and detailed study of the dynamic properties of the C-terminus of rhodopsin. The rhodopsin C-terminus is immobilized until Ala333, after which it becomes unstructured. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
| Keywords: | NMR spectroscopy Mammalian GPCR Rhodopsin Isotope labeling |
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