Molecular Characterization of Oxysterol Binding to the Epstein-Barr Virus-induced Gene 2 (GPR183)
Molecular Characterization of Oxysterol Binding to the Epstein-Barr Virus-induced Gene 2 (GPR183)
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DOI：
10.1074/jbc.m112.387894
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发表时间：
2012-10-12
影响因子：
4.8
通讯作者：
Rosenkilde, Mette M.
中科院分区：
文献类型：
--
作者：
Benned-Jensen, Tau;Norn, Christoffer;Rosenkilde, Mette M.
Oxysterols are oxygenated cholesterol derivates that are emerging as a physiologically important group of molecules. Although they regulate a range of cellular processes, only few oxysterol-binding effector proteins have been identified, and the knowledge of their binding mode is limited. Recently, the family of G protein-coupled seven transmembrane-spanning receptors (7TM receptors) was added to this group. Specifically, the Epstein-Barr virus-induced gene 2 (EBI2 or GPR183) was shown to be activated by several oxysterols, most potently by 7 alpha,25-dihydroxycholesterol (7 alpha,25-OHC). Nothing is known about the binding mode, however. Using mutational analysis, we identify here four key residues for 7 alpha,25-OHC binding: Arg-87 in TM-II (position II: 20/2.60), Tyr-112 and Tyr-116 (positions III: 09/3.33 and III: 13/3.37) in TM-III, and Tyr-260 in TM-VI (position VI: 16/6.51). Substituting these residues with Ala and/or Phe results in a severe decrease in agonist binding and receptor activation. Docking simulations suggest that Tyr-116 interacts with the 3 beta-OH group in the agonist, Tyr-260 with the 7 alpha-OH group, and Arg-87, either directly or indirectly, with the 25-OH group, although nearby residues likely also contribute. In addition, Tyr-112 is involved in 7 alpha,25-OHC binding but via hydrophobic interactions. Finally, we show that II: 20/2.60 constitutes an important residue for ligand binding in receptors carrying a positively charged residue at this position. This group is dominated by lipid-and nucleotide-activated receptors, here exemplified by the CysLTs, P2Y12, and P2Y14. In conclusion, we present the first molecular characterization of oxysterol binding to a 7TM receptor and identify position II: 20/2.60 as a generally important residue for ligand binding in certain 7TM receptors.
