+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 5kbs | ||||||
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タイトル | Cryo-EM structure of GluA2-0xSTZ at 8.7 Angstrom resolution | ||||||
要素 | Glutamate receptor 2,Voltage-dependent calcium channel gamma-2 subunit | ||||||
キーワード | TRANSPORT PROTEIN (運搬体タンパク質) / Cryo-EM (低温電子顕微鏡法) | ||||||
機能・相同性 | 機能・相同性情報 Presynaptic depolarization and calcium channel opening / LGI-ADAM interactions / regulation of postsynaptic neurotransmitter receptor activity / Trafficking of AMPA receptors / eye blink reflex / positive regulation of protein localization to basolateral plasma membrane / cerebellar mossy fiber / neurotransmitter receptor transport, postsynaptic endosome to lysosome / regulation of AMPA receptor activity / neurotransmitter receptor internalization ...Presynaptic depolarization and calcium channel opening / LGI-ADAM interactions / regulation of postsynaptic neurotransmitter receptor activity / Trafficking of AMPA receptors / eye blink reflex / positive regulation of protein localization to basolateral plasma membrane / cerebellar mossy fiber / neurotransmitter receptor transport, postsynaptic endosome to lysosome / regulation of AMPA receptor activity / neurotransmitter receptor internalization / channel regulator activity / membrane hyperpolarization / postsynaptic neurotransmitter receptor diffusion trapping / nervous system process / protein targeting to membrane / neurotransmitter receptor localization to postsynaptic specialization membrane / neuromuscular junction development / spine synapse / dendritic spine neck / dendritic spine head / Activation of AMPA receptors / voltage-gated calcium channel complex / response to lithium ion / perisynaptic space / cellular response to glycine / transmission of nerve impulse / AMPA glutamate receptor activity / regulation of postsynaptic membrane neurotransmitter receptor levels / Trafficking of GluR2-containing AMPA receptors / 脱分極 / immunoglobulin binding / AMPA glutamate receptor complex / kainate selective glutamate receptor activity / ionotropic glutamate receptor complex / extracellularly glutamate-gated ion channel activity / asymmetric synapse / regulation of receptor recycling / Unblocking of NMDA receptors, glutamate binding and activation / voltage-gated calcium channel activity / glutamate receptor binding / positive regulation of synaptic transmission / glutamate-gated receptor activity / presynaptic active zone membrane / response to fungicide / regulation of synaptic transmission, glutamatergic / cellular response to brain-derived neurotrophic factor stimulus / somatodendritic compartment / dendrite membrane / ligand-gated monoatomic ion channel activity involved in regulation of presynaptic membrane potential / ionotropic glutamate receptor binding / hippocampal mossy fiber to CA3 synapse / cytoskeletal protein binding / ionotropic glutamate receptor signaling pathway / dendrite cytoplasm / regulation of membrane potential / positive regulation of synaptic transmission, glutamatergic / SNARE binding / dendritic shaft / transmitter-gated monoatomic ion channel activity involved in regulation of postsynaptic membrane potential / synaptic membrane / synaptic transmission, glutamatergic / PDZ domain binding / postsynaptic density membrane / protein tetramerization / modulation of chemical synaptic transmission / Schaffer collateral - CA1 synapse / establishment of protein localization / terminal bouton / receptor internalization / synaptic vesicle membrane / cerebral cortex development / response to calcium ion / シナプス小胞 / presynapse / signaling receptor activity / presynaptic membrane / amyloid-beta binding / 成長円錐 / chemical synaptic transmission / perikaryon / postsynaptic membrane / scaffold protein binding / 樹状突起スパイン / postsynaptic density / neuron projection / 神経繊維 / 樹状突起 / neuronal cell body / glutamatergic synapse / シナプス / protein-containing complex binding / endoplasmic reticulum membrane / protein kinase binding / 細胞膜 / 小胞体 / protein-containing complex / 生体膜 / identical protein binding / 細胞膜 類似検索 - 分子機能 | ||||||
生物種 | Rattus norvegicus (ドブネズミ) Mus musculus (ハツカネズミ) | ||||||
手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 8.7 Å | ||||||
データ登録者 | Twomey, E.C. / Yelshanskaya, M.V. / Grassucci, R.A. / Frank, J. / Sobolevsky, A.I. | ||||||
資金援助 | 米国, 1件
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引用 | ジャーナル: Science / 年: 2016 タイトル: Elucidation of AMPA receptor-stargazin complexes by cryo-electron microscopy. 著者: Edward C Twomey / Maria V Yelshanskaya / Robert A Grassucci / Joachim Frank / Alexander I Sobolevsky / 要旨: AMPA-subtype ionotropic glutamate receptors (AMPARs) mediate fast excitatory neurotransmission and contribute to high cognitive processes such as learning and memory. In the brain, AMPAR trafficking, ...AMPA-subtype ionotropic glutamate receptors (AMPARs) mediate fast excitatory neurotransmission and contribute to high cognitive processes such as learning and memory. In the brain, AMPAR trafficking, gating, and pharmacology is tightly controlled by transmembrane AMPAR regulatory proteins (TARPs). Here, we used cryo-electron microscopy to elucidate the structural basis of AMPAR regulation by one of these auxiliary proteins, TARP γ2, or stargazin (STZ). Our structures illuminate the variable interaction stoichiometry of the AMPAR-TARP complex, with one or two TARP molecules binding one tetrameric AMPAR. Analysis of the AMPAR-STZ binding interfaces suggests that electrostatic interactions between the extracellular domains of AMPAR and STZ play an important role in modulating AMPAR function through contact surfaces that are conserved across AMPARs and TARPs. We propose a model explaining how TARPs stabilize the activated state of AMPARs and how the interactions between AMPARs and their auxiliary proteins control fast excitatory synaptic transmission. | ||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | 分子: MolmilJmol/JSmol |
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 5kbs.cif.gz | 593.9 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb5kbs.ent.gz | 464.2 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 5kbs.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
その他 | その他のダウンロード |
-検証レポート
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/kb/5kbs ftp://data.pdbj.org/pub/pdb/validation_reports/kb/5kbs | HTTPS FTP |
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-関連構造データ
-リンク
-集合体
登録構造単位 |
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-要素
#1: タンパク質 | 分子量: 115515.984 Da / 分子数: 4 変異: N241E, V382L, G384E, N385D, V758L,N241E, V382L, G384E, N385D, V758L 由来タイプ: 組換発現 由来: (組換発現) Rattus norvegicus (ドブネズミ), (組換発現) Mus musculus (ハツカネズミ) 遺伝子: Gria2, Glur2, Cacng2, Stg / 発現宿主: Homo sapiens (ヒト) / 参照: UniProt: P19491, UniProt: O88602 #2: 化合物 | ChemComp-ZK1 / {[ #3: 糖 | ChemComp-NAG / 構成要素の詳細 | The protein was expressed as a tandem fusion construct (GluA2 fused to stargazin, also named TARP- ...The protein was expressed as a tandem fusion construct (GluA2 fused to stargazin, also named TARP-y2 or cagcn2). However, this second protein which is covalently linked or fused to the C-terminus of GluA2 is not observed in this structure. | |
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-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法 |
-試料調製
構成要素 | 名称: Proteinタンパク質 / タイプ: COMPLEX / Entity ID: #1-#2 / 由来: RECOMBINANT |
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由来(天然) | 生物種: Rattus norvegicus (ドブネズミ) |
由来(組換発現) | 生物種: Homo sapiens (ヒト) / 細胞: HEK293 / プラスミド: Bacmam |
緩衝液 | pH: 8 |
試料 | 濃度: 2 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
試料支持 | 詳細: The grid was coated with gold prior to use. / グリッドの材料: GOLD / グリッドのサイズ: 200 divisions/in. / グリッドのタイプ: C-Flat Au 1.2/1.3 |
急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 295 K / 詳細: 3 blot force, 8.0 s blot time |
-電子顕微鏡撮影
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TITAN KRIOS |
電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: SPOT SCAN |
電子レンズ | モード: BRIGHT FIELDBright-field microscopy |
試料ホルダ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
撮影 | 平均露光時間: 8 sec. / 電子線照射量: 80 e/Å2 / 検出モード: SUPER-RESOLUTION フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 撮影したグリッド数: 1 詳細: 40 frames were collected across 8 seconds per image. |
画像スキャン | 動画フレーム数/画像: 40 / 利用したフレーム数/画像: 1-40 |
-解析
EMソフトウェア |
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CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||
対称性 | 点対称性: C2 (2回回転対称) | ||||||||||||||||||||
3次元再構成 | 解像度: 8.7 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 7993 / 対称性のタイプ: POINT |