Isotope labeling of mammalian GPCRs in HEK293 cells and characterization of the C-terminus of bovine rhodopsin by high resolution liquid NMR spectroscopy.
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.
Werner
Karla
K
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.
Richter
Christian
C
Klein-Seetharaman
Judith
J
Schwalbe
Harald
H
D000595:Amino Acid Sequence; D000818:Animals; D002247:Carbon Isotopes; D002417:Cattle; D002460:Cell Line; D006801:Humans; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D012243:Rhodopsin / Q000737:chemistry* / Q000378:metabolism
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D012243:Rhodopsin