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.
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Acknowledgments
We thank for the financial support from the Sofya Kovalevskaya-Award granted by the Alexander von Humboldt-Foundation through the Investment in the Future Programme of the German Federal Government. The work was supported by DFG (SFB 628: “Functional Membrane Proteomics”), the State of Hesse (BMRZ) and the Fonds der Chemischen Industrie (H.S.). We acknowledge support by Cambridge Isotope Laboratories with isotope labeled material.
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Werner, K., Richter, C., Klein-Seetharaman, J. et al. Isotope labeling of mammalian GPCRs in HEK293 cells and characterization of the C-terminus of bovine rhodopsin by high resolution liquid NMR spectroscopy. J Biomol NMR 40, 49–53 (2008). https://doi.org/10.1007/s10858-007-9205-3
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DOI: https://doi.org/10.1007/s10858-007-9205-3