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基本情報
登録情報 | データベース: EMDB / ID: EMD-21685 | |||||||||||||||||||||
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タイトル | Structure of human GABA(B) receptor in an inactive state | |||||||||||||||||||||
![]() | Final composite map combining local reconstructions for the extracellular and transmembrane domains. Map was re-sampled in Chimera with pixel spacing 1.1 angstrom. | |||||||||||||||||||||
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![]() | G protein coupled receptor (GPCR) / GABA / GABA(B) receptor / Class C GPCR / Phospholipids / Inhibitory neurotransmission. / MEMBRANE PROTEIN | |||||||||||||||||||||
機能・相同性 | ![]() negative regulation of gamma-aminobutyric acid secretion / GABA B receptor activation / G protein-coupled GABA receptor complex / G protein-coupled neurotransmitter receptor activity involved in regulation of postsynaptic membrane potential / neuron-glial cell signaling / G protein-coupled neurotransmitter receptor activity involved in regulation of presynaptic membrane potential / negative regulation of dopamine secretion / G protein-coupled GABA receptor activity / G protein-coupled receptor heterodimeric complex / negative regulation of epinephrine secretion ...negative regulation of gamma-aminobutyric acid secretion / GABA B receptor activation / G protein-coupled GABA receptor complex / G protein-coupled neurotransmitter receptor activity involved in regulation of postsynaptic membrane potential / neuron-glial cell signaling / G protein-coupled neurotransmitter receptor activity involved in regulation of presynaptic membrane potential / negative regulation of dopamine secretion / G protein-coupled GABA receptor activity / G protein-coupled receptor heterodimeric complex / negative regulation of epinephrine secretion / positive regulation of growth hormone secretion / extracellular matrix protein binding / GABA receptor complex / negative regulation of adenylate cyclase activity / Class C/3 (Metabotropic glutamate/pheromone receptors) / negative regulation of synaptic transmission / positive regulation of glutamate secretion / gamma-aminobutyric acid signaling pathway / synaptic transmission, GABAergic / axolemma / dendritic shaft / response to nicotine / adenylate cyclase-inhibiting G protein-coupled receptor signaling pathway / mitochondrial membrane / GABA-ergic synapse / Schaffer collateral - CA1 synapse / Activation of G protein gated Potassium channels / Inhibition of voltage gated Ca2+ channels via Gbeta/gamma subunits / osteoblast differentiation / synaptic vesicle / transmembrane signaling receptor activity / presynaptic membrane / response to ethanol / G alpha (i) signalling events / chemical synaptic transmission / dendritic spine / postsynaptic membrane / neuron projection / G protein-coupled receptor signaling pathway / protein heterodimerization activity / negative regulation of cell population proliferation / neuronal cell body / endoplasmic reticulum membrane / glutamatergic synapse / extracellular space / plasma membrane / cytoplasm 類似検索 - 分子機能 | |||||||||||||||||||||
生物種 | ![]() | |||||||||||||||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.3 Å | |||||||||||||||||||||
![]() | Park J / Fu Z / Frangaj A / Liu J / Mosyak L / Shen T / Slavkovich VN / Ray KM / Taura J / Cao B ...Park J / Fu Z / Frangaj A / Liu J / Mosyak L / Shen T / Slavkovich VN / Ray KM / Taura J / Cao B / Geng Y / Zuo H / Kou Y / Grassucci R / Chen S / Liu Z / Lin X / Williams JP / Rice WJ / Eng ET / Huang RK / Soni RK / Kloss B / Yu Z / Javitch JA / Hendrickson WA / Slesinger PA / Quick M / Graziano J / Yu H / Fiehn O / Clarke OB / Frank J / Fan QR | |||||||||||||||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Structure of human GABA receptor in an inactive state. 著者: Jinseo Park / Ziao Fu / Aurel Frangaj / Jonathan Liu / Lidia Mosyak / Tong Shen / Vesna N Slavkovich / Kimberly M Ray / Jaume Taura / Baohua Cao / Yong Geng / Hao Zuo / Yongjun Kou / Robert ...著者: Jinseo Park / Ziao Fu / Aurel Frangaj / Jonathan Liu / Lidia Mosyak / Tong Shen / Vesna N Slavkovich / Kimberly M Ray / Jaume Taura / Baohua Cao / Yong Geng / Hao Zuo / Yongjun Kou / Robert Grassucci / Shaoxia Chen / Zheng Liu / Xin Lin / Justin P Williams / William J Rice / Edward T Eng / Rick K Huang / Rajesh K Soni / Brian Kloss / Zhiheng Yu / Jonathan A Javitch / Wayne A Hendrickson / Paul A Slesinger / Matthias Quick / Joseph Graziano / Hongtao Yu / Oliver Fiehn / Oliver B Clarke / Joachim Frank / Qing R Fan / ![]() ![]() ![]() 要旨: The human GABA receptor-a member of the class C family of G-protein-coupled receptors (GPCRs)-mediates inhibitory neurotransmission and has been implicated in epilepsy, pain and addiction. A unique ...The human GABA receptor-a member of the class C family of G-protein-coupled receptors (GPCRs)-mediates inhibitory neurotransmission and has been implicated in epilepsy, pain and addiction. A unique GPCR that is known to require heterodimerization for function, the GABA receptor has two subunits, GABA and GABA, that are structurally homologous but perform distinct and complementary functions. GABA recognizes orthosteric ligands, while GABA couples with G proteins. Each subunit is characterized by an extracellular Venus flytrap (VFT) module, a descending peptide linker, a seven-helix transmembrane domain and a cytoplasmic tail. Although the VFT heterodimer structure has been resolved, the structure of the full-length receptor and its transmembrane signalling mechanism remain unknown. Here we present a near full-length structure of the GABA receptor, captured in an inactive state by cryo-electron microscopy. Our structure reveals several ligands that preassociate with the receptor, including two large endogenous phospholipids that are embedded within the transmembrane domains to maintain receptor integrity and modulate receptor function. We also identify a previously unknown heterodimer interface between transmembrane helices 3 and 5 of both subunits, which serves as a signature of the inactive conformation. A unique 'intersubunit latch' within this transmembrane interface maintains the inactive state, and its disruption leads to constitutive receptor activity. | |||||||||||||||||||||
履歴 |
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構造の表示
ムービー |
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構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
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ダウンロードとリンク
-EMDBアーカイブ
マップデータ | ![]() | 45.8 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 30.8 KB 30.8 KB | 表示 表示 | ![]() |
画像 | ![]() | 146.5 KB | ||
Filedesc metadata | ![]() | 8.6 KB | ||
その他 | ![]() ![]() ![]() | 47.6 MB 47.9 MB 47.4 MB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 474.2 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 473.8 KB | 表示 | |
XML形式データ | ![]() | 5.9 KB | 表示 | |
CIF形式データ | ![]() | 6.8 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 6wivMC M: このマップから作成された原子モデル C: 同じ文献を引用 ( |
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類似構造データ | |
電子顕微鏡画像生データ | ![]() Data size: 854.1 Data #1: Unaligned multi-frame micrographs of GABA(B) receptor in the inactive state [micrographs - multiframe]) |
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リンク
EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Final composite map combining local reconstructions for the extracellular and transmembrane domains. Map was re-sampled in Chimera with pixel spacing 1.1 angstrom. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.1 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
-追加マップ: Local reconstruction for the extracellular domain. Map was...
ファイル | emd_21685_additional_1.map | ||||||||||||
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注釈 | Local reconstruction for the extracellular domain. Map was re-sampled in Chimera with pixel spacing 1.1 angstrom. | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Local reconstruction for the transmembrane domain. Map was...
ファイル | emd_21685_additional_2.map | ||||||||||||
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注釈 | Local reconstruction for the transmembrane domain. Map was re-sampled in Chimera with pixel spacing 1.1 angstrom. | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Global map without local refinement. Map was re-sampled...
ファイル | emd_21685_additional_3.map | ||||||||||||
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注釈 | Global map without local refinement. Map was re-sampled in Chimera with pixel spacing 1.1 angstrom. | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
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試料の構成要素
-全体 : Heterodimer human GABA(B) receptor composed of GABA(B1b) and GABA...
全体 | 名称: Heterodimer human GABA(B) receptor composed of GABA(B1b) and GABA(B2) subunits. |
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要素 |
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-超分子 #1: Heterodimer human GABA(B) receptor composed of GABA(B1b) and GABA...
超分子 | 名称: Heterodimer human GABA(B) receptor composed of GABA(B1b) and GABA(B2) subunits. タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#2 |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 193 KDa |
-分子 #1: Gamma-aminobutyric acid type B receptor subunit 1
分子 | 名称: Gamma-aminobutyric acid type B receptor subunit 1 / タイプ: protein_or_peptide / ID: 1 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 91.585867 KDa |
組換発現 | 生物種: ![]() |
配列 | 文字列: MGPGAPFARV GWPLPLLVVM AAGVAPVWAS HSPHLPRPHS RVPPHPSSER RAVYIGALFP MSGGWPGGQA CQPAVEMALE DVNSRRDIL PDYELKLIHH DSKCDPGQAT KYLYELLYND PIKIILMPGC SSVSTLVAEA ARMWNLIVLS YGSSSPALSN R QRFPTFFR ...文字列: MGPGAPFARV GWPLPLLVVM AAGVAPVWAS HSPHLPRPHS RVPPHPSSER RAVYIGALFP MSGGWPGGQA CQPAVEMALE DVNSRRDIL PDYELKLIHH DSKCDPGQAT KYLYELLYND PIKIILMPGC SSVSTLVAEA ARMWNLIVLS YGSSSPALSN R QRFPTFFR THPSATLHNP TRVKLFEKWG WKKIATIQQT TEVFTSTLDD LEERVKEAGI EITFRQSFFS DPAVPVKNLK RQ DARIIVG LFYETEARKV FCEVYKERLF GKKYVWFLIG WYADNWFKIY DPSINCTVDE MTEAVEGHIT TEIVMLNPAN TRS ISNMTS QEFVEKLTKR LKRHPEETGG FQEAPLAYDA IWALALALNK TSGGGGRSGV RLEDFNYNNQ TITDQIYRAM NSSS FEGVS GHVVFDASGS RMAWTLIEQL QGGSYKKIGY YDSTKDDLSW SKTDKWIGGS PPADQTLVIK TFRFLSQKLF ISVSV LSSL GIVLAVVCLS FNIYNSHVRY IQNSQPNLNN LTAVGCSLAL AAVFPLGLDG YHIGRNQFPF VCQARLWLLG LGFSLG YGS MFTKIWWVHT VFTKKEEKKE WRKTLEPWKL YATVGLLVGM DVLTLAIWQI VDPLHRTIET FAKEEPKEDI DVSILPQ LE HCSSRKMNTW LGIFYGYKGL LLLLGIFLAY ETKSVSTEKI NDHRAVGMAI YNVAVLCLIT APVTMILSSQ QDAAFAFA S LAIVFSSYIT LVVLFVPKMR RLITRGEWQS EAQDTMKTGS STNNNEEEKS RLLEKENREL EKIIAEKEER VSELRHQLQ SRDYKDDDDK UniProtKB: Gamma-aminobutyric acid type B receptor subunit 1 |
-分子 #2: Gamma-aminobutyric acid type B receptor subunit 2
分子 | 名称: Gamma-aminobutyric acid type B receptor subunit 2 / タイプ: protein_or_peptide / ID: 2 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 93.278781 KDa |
組換発現 | 生物種: ![]() |
配列 | 文字列: MASPRSSGQP GPPPPPPPPP ARLLLLLLLP LLLPLAPGAW GWARGAPRPP PSSPPLSIMG LMPLTKEVAK GSIGRGVLPA VELAIEQIR NESLLRPYFL DLRLYDTECD NAKGLKAFYD AIKYGPNHLM VFGGVCPSVT SIIAESLQGW NLVQLSFAAT T PVLADKKK ...文字列: MASPRSSGQP GPPPPPPPPP ARLLLLLLLP LLLPLAPGAW GWARGAPRPP PSSPPLSIMG LMPLTKEVAK GSIGRGVLPA VELAIEQIR NESLLRPYFL DLRLYDTECD NAKGLKAFYD AIKYGPNHLM VFGGVCPSVT SIIAESLQGW NLVQLSFAAT T PVLADKKK YPYFFRTVPS DNAVNPAILK LLKHYQWKRV GTLTQDVQRF SEVRNDLTGV LYGEDIEISD TESFSNDPCT SV KKLKGND VRIILGQFDQ NMAAKVFCCA YEENMYGSKY QWIIPGWYEP SWWEQVHTEA NSSRCLRKNL LAAMEGYIGV DFE PLSSKQ IKTISGKTPQ QYEREYNNKR SGVGPSKFHG YAYDGIWVIA KTLQRAMETL HASSRHQRIQ DFNYTDHTLG RIIL NAMNE TNFFGVTGQV VFRNGERMGT IKFTQFQDSR EVKVGEYNAV ADTLEIINDT IRFQGSEPPK DKTIILEQLR KISLP LYSI LSALTILGMI MASAFLFFNI KNRNQKLIKM SSPYMNNLII LGGMLSYASI FLFGLDGSFV SEKTFETLCT VRTWIL TVG YTTAFGAMFA KTWRVHAIFK NVKMKKKIIK DQKLLVIVGG MLLIDLCILI CWQAVDPLRR TVEKYSMEPD PAGRDIS IR PLLEHCENTH MTIWLGIVYA YKGLLMLFGC FLAWETRNVS IPALNDSKYI GMSVYNVGIM CIIGAAVSFL TRDQPNVQ F CIVALVIIFC STITLCLVFV PKLITLRTNP DAATQNRRFQ FTQNQKKEDS KTSTSVTSVN QASTSRLEGL QSENHRLRM KITELDKDLE EVTMQLQDTD YKDDDDK UniProtKB: Gamma-aminobutyric acid type B receptor subunit 2 |
-分子 #3: 2-acetamido-2-deoxy-beta-D-glucopyranose
分子 | 名称: 2-acetamido-2-deoxy-beta-D-glucopyranose / タイプ: ligand / ID: 3 / コピー数: 4 / 式: NAG |
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分子量 | 理論値: 221.208 Da |
Chemical component information | ![]() ChemComp-NAG: |
-分子 #4: CALCIUM ION
分子 | 名称: CALCIUM ION / タイプ: ligand / ID: 4 / コピー数: 1 / 式: CA |
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分子量 | 理論値: 40.078 Da |
-分子 #5: (2R)-3-{[(S)-(2-aminoethoxy)(hydroxy)phosphoryl]oxy}-2-{[(9Z)-oct...
分子 | 名称: (2R)-3-{[(S)-(2-aminoethoxy)(hydroxy)phosphoryl]oxy}-2-{[(9Z)-octadec-9-enoyl]oxy}propyl (5Z,8Z,11Z,14Z)-icosa-5,8,11,14-tetraenoate タイプ: ligand / ID: 5 / コピー数: 1 / 式: U3G |
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分子量 | 理論値: 766.039 Da |
Chemical component information | ![]() ChemComp-U3G: |
-分子 #6: CHOLESTEROL
分子 | 名称: CHOLESTEROL / タイプ: ligand / ID: 6 / コピー数: 10 / 式: CLR |
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分子量 | 理論値: 386.654 Da |
Chemical component information | ![]() ChemComp-CLR: |
-分子 #7: [(2R)-3-[(Z)-icos-11-enoyl]oxy-2-[(Z)-octadec-9-enoyl]oxypropyl] ...
分子 | 名称: [(2R)-3-[(Z)-icos-11-enoyl]oxy-2-[(Z)-octadec-9-enoyl]oxypropyl] 2-(trimethylazaniumyl)ethyl phosphate タイプ: ligand / ID: 7 / コピー数: 1 / 式: U3D |
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分子量 | 理論値: 814.167 Da |
Chemical component information | ![]() ChemComp-U3D: |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
濃度 | 0.3 mg/mL | |||||||||||||||
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緩衝液 | pH: 7.5 構成要素:
詳細: Solutions were made fresh from concentrated and filtered to avoid microbial contamination. | |||||||||||||||
グリッド | モデル: Quantifoil R0.6/1 / 材質: GOLD / メッシュ: 300 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY / 支持フィルム - Film thickness: 50 / 前処理 - タイプ: PLASMA CLEANING / 前処理 - 時間: 25 sec. / 前処理 - 雰囲気: OTHER | |||||||||||||||
凍結 | 凍結剤: ETHANE-PROPANE / チャンバー内湿度: 100 % / チャンバー内温度: 277 K / 装置: FEI VITROBOT MARK IV / 詳細: blot for 4 seconds before plunging. | |||||||||||||||
詳細 | This sample was monodisperse |
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電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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温度 | 最高: 100.0 K |
特殊光学系 | エネルギーフィルター - スリット幅: 20 eV |
詳細 | Preliminary grid screening was performed manually. |
撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: COUNTING / デジタル化 - 画像ごとのフレーム数: 1-60 / 撮影したグリッド数: 1 / 実像数: 3435 / 平均露光時間: 12.0 sec. / 平均電子線量: 85.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | C2レンズ絞り径: 70.0 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm / 最大 デフォーカス(公称値): -2.0 µm / 最小 デフォーカス(公称値): -0.5 µm / 倍率(公称値): 130000 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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画像解析
-原子モデル構築 1
初期モデル |
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精密化 | 空間: REAL / プロトコル: FLEXIBLE FIT | ||||||
得られたモデル | ![]() PDB-6wiv: |