ジャーナル: Nat Commun / 年: 2021 タイトル: Structures of active-state orexin receptor 2 rationalize peptide and small-molecule agonist recognition and receptor activation. 著者: Chuan Hong / Noel J Byrne / Beata Zamlynny / Srivanya Tummala / Li Xiao / Jennifer M Shipman / Andrea T Partridge / Christina Minnick / Michael J Breslin / Michael T Rudd / Shawn J Stachel / ...著者: Chuan Hong / Noel J Byrne / Beata Zamlynny / Srivanya Tummala / Li Xiao / Jennifer M Shipman / Andrea T Partridge / Christina Minnick / Michael J Breslin / Michael T Rudd / Shawn J Stachel / Vanessa L Rada / Jeffrey C Kern / Kira A Armacost / Scott A Hollingsworth / Julie A O'Brien / Dawn L Hall / Terrence P McDonald / Corey Strickland / Alexei Brooun / Stephen M Soisson / Kaspar Hollenstein / 要旨: Narcolepsy type 1 (NT1) is a chronic neurological disorder that impairs the brain's ability to control sleep-wake cycles. Current therapies are limited to the management of symptoms with modest ...Narcolepsy type 1 (NT1) is a chronic neurological disorder that impairs the brain's ability to control sleep-wake cycles. Current therapies are limited to the management of symptoms with modest effectiveness and substantial adverse effects. Agonists of the orexin receptor 2 (OXR) have shown promise as novel therapeutics that directly target the pathophysiology of the disease. However, identification of drug-like OXR agonists has proven difficult. Here we report cryo-electron microscopy structures of active-state OXR bound to an endogenous peptide agonist and a small-molecule agonist. The extended carboxy-terminal segment of the peptide reaches into the core of OXR to stabilize an active conformation, while the small-molecule agonist binds deep inside the orthosteric pocket, making similar key interactions. Comparison with antagonist-bound OXR suggests a molecular mechanism that rationalizes both receptor activation and inhibition. Our results enable structure-based discovery of therapeutic orexin agonists for the treatment of NT1 and other hypersomnia disorders.
凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER 最高温度: 93 K / 最低温度: 83 K
撮影
平均露光時間: 0.05 sec. / 電子線照射量: 1.0625 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 撮影したグリッド数: 2 / 実像数: 18000 / 詳細: 40 frames with total 2s exposure per movie
電子光学装置
エネルギーフィルター名称: GIF Bioquantum / エネルギーフィルタースリット幅: 20 eV
画像スキャン
サンプリングサイズ: 5 µm / 横: 5700 / 縦: 4100
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解析
EMソフトウェア
ID
名称
バージョン
カテゴリ
詳細
1
cryoSPARC
2.15
粒子像選択
2
Latitude
3.32.2403
画像取得
Gatan
4
cryoSPARC
2.15
CTF補正
7
Coot
0.8.9.2
モデルフィッティング
9
cryoSPARC
2.15
初期オイラー角割当
10
cryoSPARC
2.15
最終オイラー角割当
11
cryoSPARC
2.15
分類
12
cryoSPARC
2.15
3次元再構成
13
PHENIX
1.15
モデル精密化
CTF補正
タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION
粒子像の選択
選択した粒子像数: 7500000
対称性
点対称性: C1 (非対称)
3次元再構成
解像度: 3 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 1100000 / アルゴリズム: FOURIER SPACE / 詳細: Gold standard with masking effect correction / クラス平均像の数: 1 / 対称性のタイプ: POINT