National Institutes of Health/National Heart, Lung, and Blood Institute
HL16037
米国
National Institutes of Health/National Institute of Neurological Disorders and Stroke
NS092695
米国
引用
ジャーナル: Nature / 年: 2020 タイトル: Structure of the M2 muscarinic receptor-β-arrestin complex in a lipid nanodisc. 著者: Dean P Staus / Hongli Hu / Michael J Robertson / Alissa L W Kleinhenz / Laura M Wingler / William D Capel / Naomi R Latorraca / Robert J Lefkowitz / Georgios Skiniotis / 要旨: After activation by an agonist, G-protein-coupled receptors (GPCRs) recruit β-arrestin, which desensitizes heterotrimeric G-protein signalling and promotes receptor endocytosis. Additionally, β- ...After activation by an agonist, G-protein-coupled receptors (GPCRs) recruit β-arrestin, which desensitizes heterotrimeric G-protein signalling and promotes receptor endocytosis. Additionally, β-arrestin directly regulates many cell signalling pathways that can induce cellular responses distinct from that of G proteins. In contrast to G proteins, for which there are many high-resolution structures in complex with GPCRs, the molecular mechanisms underlying the interaction of β-arrestin with GPCRs are much less understood. Here we present a cryo-electron microscopy structure of β-arrestin 1 (βarr1) in complex with M2 muscarinic receptor (M2R) reconstituted in lipid nanodiscs. The M2R-βarr1 complex displays a multimodal network of flexible interactions, including binding of the N domain of βarr1 to phosphorylated receptor residues and insertion of the finger loop of βarr1 into the M2R seven-transmembrane bundle, which adopts a conformation similar to that in the M2R-heterotrimeric G protein complex. Moreover, the cryo-electron microscopy map reveals that the C-edge of βarr1 engages the lipid bilayer. Through atomistic simulations and biophysical, biochemical and cellular assays, we show that the C-edge is critical for stable complex formation, βarr1 recruitment, receptor internalization, and desensitization of G-protein activation. Taken together, these data suggest that the cooperative interactions of β-arrestin with both the receptor and the phospholipid bilayer contribute to its functional versatility.
ダウンロード / ファイル: emd_20948.map.gz / 形式: CCP4 / 大きさ: 52.7 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
GPCR-Beta arrestin in lipid bilayer
ボクセルのサイズ
X=Y=Z: 1.06 Å
密度
表面レベル
登録者による: 4.2 / ムービー #1: 4.2
最小 - 最大
-13.553397 - 30.015896
平均 (標準偏差)
0.0070778565 (±0.83977336)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
0
0
0
サイズ
240
240
240
Spacing
240
240
240
セル
A=B=C: 254.4 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
1.06
1.06
1.06
M x/y/z
240
240
240
origin x/y/z
0.000
0.000
0.000
length x/y/z
254.400
254.400
254.400
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
240
240
240
D min/max/mean
-13.553
30.016
0.007
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添付データ
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試料の構成要素
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全体 : Phosphorylated human muscarinic acetylcholine receptor M2 in comp...
全体
名称: Phosphorylated human muscarinic acetylcholine receptor M2 in complex with rat beta-arrestin1, stabilized by an antibody fragment (Fab30).
要素
複合体: Phosphorylated human muscarinic acetylcholine receptor M2 in complex with rat beta-arrestin1, stabilized by an antibody fragment (Fab30).
複合体: Phosphorylated human muscarinic acetylcholine receptor M2
複合体: Cysteine-free rat beta-arrestin 1 truncated at amino acid 393
複合体: Antibody Fragment (Fab30)
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超分子 #1: Phosphorylated human muscarinic acetylcholine receptor M2 in comp...
超分子
名称: Phosphorylated human muscarinic acetylcholine receptor M2 in complex with rat beta-arrestin1, stabilized by an antibody fragment (Fab30). タイプ: complex / ID: 1 / 親要素: 0
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超分子 #2: Phosphorylated human muscarinic acetylcholine receptor M2
超分子
名称: Phosphorylated human muscarinic acetylcholine receptor M2 タイプ: complex / ID: 2 / 親要素: 1 詳細: Phosphorylated M2R was generated by ligating a synthetic phosphopeptide derived from the vasopressin-2-receptor (V2Rpp) using the enzyme sortase.
由来(天然)
生物種: Homo sapiens (ヒト)
組換発現
生物種: Homo sapiens (ヒト)
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超分子 #3: Cysteine-free rat beta-arrestin 1 truncated at amino acid 393
超分子
名称: Cysteine-free rat beta-arrestin 1 truncated at amino acid 393 タイプ: complex / ID: 3 / 親要素: 1