Research ArticleSTRUCTURAL BIOLOGY

Structure-based mechanism of cysteinyl leukotriene receptor inhibition by antiasthmatic drugs

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Science Advances  09 Oct 2019:
Vol. 5, no. 10, eaax2518
DOI: 10.1126/sciadv.aax2518
  • Fig. 1 Overall structure of CysLT1R and its comparison with other receptors.

    (A) Side and (B) top views of CysLT1R-pran (receptor, orange; ligand, blue) and CysLT1R-zafir (receptor, green; ligand, yellow). (C) Superposition of CysLT1R-pran with BLT1-BIIL260 [Protein Data Bank (PDB) ID 5X33, light blue] and P2Y1R-BPTU (PDB ID 4XNV, pink). (D) Distribution of lipid receptors on the human GPCR sequence homology tree. CysLT1R and CysLT2R are marked as red dots, and other lipid receptors are marked as blue dots. The percentage of the lipid receptors located on each branch is shown, with the exact numbers given in parenthesis. The membrane boundaries are shown as dashed lines in (A) and (C).

  • Fig. 2 Functional motifs of CysLT1R show unusual inactive-state features.

    (A) Superposition of CysLT1R-pran (orange) with β2AR in inactive (PDB ID 2RH1; violet) and active (PDB ID 3SN6; teal) conformations. The membrane boundaries are shown as dashed lines. Loops are removed for clarity. (B to E) Zoom in on functional elements: DRY motif (B), intracellular region (C), P-I-F motif (D), and NPxxY motif (E). A different conformation of R1213.50 in CysLT1R-zafir (chain A) is shown as green sticks in (B).

  • Fig. 3 Sodium-binding pocket in CysLT1R.

    (A) Details of Na+ (purple sphere) coordination. Water molecule is shown as a red sphere. (B) Comparison of all high-resolution GPCR structures with resolved Na+. Sodium ions are shown as purple spheres for the δ-branch receptors: CysLT1R-zafir (green), PAR1 (light purple; PDB ID 3VW7), PAR2 (PDB ID 5NDD), and as yellow spheres for receptors from the α-branch: A2AAR (yellow; PDB ID 4EIY) and β1AR (turkey; PDB ID 4BVN), and the γ-branch: DOR (PDB ID 4N6H). (C and D) Frequency analysis of amino acid occurrence in the sodium pocket of the δ-branch class A GPCRs (C) and other class A receptors excluding the δ-branch (D). Yellow color marks amino acids with hydrophobic side chains; green, aromatic; red, negatively charged; blue, positively charged; purple, polar uncharged; pink, Gly and Pro. Frames indicate positions with the largest differences. The frequency analysis was performed using the weblogo.berkeley.edu server.

  • Fig. 4 Orthosteric ligand-binding pocket in CysLT1R.

    (A and D) Details of ligand-receptor interactions for pranlukast (A) and zafirlukast (D). (B and E) Pocket shapes for pranlukast (B) and zafirlukast (E). (C and F) Pocket entrance for pranlukast, closed “gate” (C), and zafirlukast, open “gate” (F). (G and H) 2D representations of receptor-ligand interactions for pranlukast (G) and zafirlukast (H). Water molecules are shown as red spheres in (A). Residues engaged in the same type of interactions with both zafirlukast and pranlukast are colored in light green, and those engaged in different types are colored in orange (G and H). The membrane boundary is shown as a dashed line in (B, C, E, and F).

Supplementary Materials

  • Supplementary material for this article is available at http://advances.sciencemag.org/cgi/content/full/5/10/eaax2518/DC1

    Supplementary Materials and Methods

    Fig. S1. Protein purification and crystallization.

    Fig. S2. Crystal packing.

    Fig. S3. Examples of electron density.

    Fig. S4. Comparison of CysLT1R-pran structure and ligand binding pose with other lipid and δ-branch class A GPCRs.

    Fig. S5. Effect of monovalent cations on CysLT1R stability.

    Fig. S6. Effects of LTD4 and CysLT1R antagonists on IP1 production.

    Fig. S7. Docking of LTD4 in CysLT1R.

    Fig. S8. MD simulations of the TM4-TM5 gate closure.

    Table S1. Crystallographic data collection and refinement statistics.

    Table S2. Signaling and cell surface expression data for CysLT1R.

    Movie S1. Rapid closure of the ligand access gate.

    Movie S2. Lipid molecule enters the ligand access gate.

    Movie S3. Spontaneous opening and closing of the ligand access gate.

    Reference (55)

  • Supplementary Materials

    The PDF file includes:

    • Supplementary Materials and Methods
    • Fig. S1. Protein purification and crystallization.
    • Fig. S2. Crystal packing.
    • Fig. S3. Examples of electron density.
    • Fig. S4. Comparison of CysLT1R-pran structure and ligand binding pose with other lipid and δ-branch class A GPCRs.
    • Fig. S5. Effect of monovalent cations on CysLT1R stability.
    • Fig. S6. Effects of LTD4 and CysLT1R antagonists on IP1 production.
    • Fig. S7. Docking of LTD4 in CysLT1R.
    • Fig. S8. MD simulations of the TM4-TM5 gate closure.
    • Table S1. Crystallographic data collection and refinement statistics.
    • Table S2. Signaling and cell surface expression data for CysLT1R.
    • Legends for movies S1 to S3
    • Reference (55)

    Download PDF

    Other Supplementary Material for this manuscript includes the following:

    • Movie S1 (.mp4 format). Rapid closure of the ligand access gate.
    • Movie S2 (.mp4 format). Lipid molecule enters the ligand access gate.
    • Movie S3 (.mp4 format). Spontaneous opening and closing of the ligand access gate.

    Files in this Data Supplement:

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