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基本情報
登録情報 | データベース: EMDB / ID: EMD-5981 | |||||||||
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タイトル | Cryo-Electron Microscopy Reconstruction of Glucocorticoid Receptor-bound Hsp90-Hsp70-Hop Chaperone Complex | |||||||||
![]() | Reconstruction of client-bound Hsp90-Hsp70-Hop chaperone complex | |||||||||
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![]() | Hsp90 / Hsp70 / Hop / Glucocorticoid Receptor | |||||||||
機能・相同性 | ![]() : / Regulation of NPAS4 gene transcription / cellular heat acclimation / regulation of glucocorticoid biosynthetic process / nuclear glucocorticoid receptor activity / negative regulation of inclusion body assembly / : / positive regulation of nucleotide-binding oligomerization domain containing 2 signaling pathway / C3HC4-type RING finger domain binding / steroid hormone binding ...: / Regulation of NPAS4 gene transcription / cellular heat acclimation / regulation of glucocorticoid biosynthetic process / nuclear glucocorticoid receptor activity / negative regulation of inclusion body assembly / : / positive regulation of nucleotide-binding oligomerization domain containing 2 signaling pathway / C3HC4-type RING finger domain binding / steroid hormone binding / positive regulation of microtubule nucleation / PTK6 Expression / ATP-dependent protein disaggregase activity / dynein axonemal particle / neuroinflammatory response / glucocorticoid metabolic process / microglia differentiation / misfolded protein binding / mammary gland duct morphogenesis / maternal behavior / regulation of mitotic spindle assembly / positive regulation of tumor necrosis factor-mediated signaling pathway / astrocyte differentiation / cellular response to interleukin-7 / cellular response to glucocorticoid stimulus / aggresome / protein folding chaperone complex / RND1 GTPase cycle / motor behavior / positive regulation of tau-protein kinase activity / sperm mitochondrial sheath / dATP binding / Scavenging by Class F Receptors / sulfonylurea receptor binding / CTP binding / positive regulation of protein polymerization / vRNP Assembly / UTP binding / sperm plasma membrane / chaperone-mediated autophagy / telomerase holoenzyme complex assembly / adrenal gland development / protein insertion into mitochondrial outer membrane / cellular response to steroid hormone stimulus / regulation of gluconeogenesis / Respiratory syncytial virus genome replication / Rho GDP-dissociation inhibitor binding / Uptake and function of diphtheria toxin / mitochondrial transport / Drug-mediated inhibition of ERBB2 signaling / Resistance of ERBB2 KD mutants to trastuzumab / Resistance of ERBB2 KD mutants to sapitinib / Resistance of ERBB2 KD mutants to tesevatinib / Resistance of ERBB2 KD mutants to neratinib / Resistance of ERBB2 KD mutants to osimertinib / Resistance of ERBB2 KD mutants to afatinib / Resistance of ERBB2 KD mutants to AEE788 / Resistance of ERBB2 KD mutants to lapatinib / Drug resistance in ERBB2 TMD/JMD mutants / PIWI-interacting RNA (piRNA) biogenesis / TPR domain binding / non-chaperonin molecular chaperone ATPase / Assembly and release of respiratory syncytial virus (RSV) virions / dendritic growth cone / mRNA catabolic process / regulation of postsynaptic membrane neurotransmitter receptor levels / Sema3A PAK dependent Axon repulsion / regulation of protein ubiquitination / skeletal muscle contraction / chaperone cofactor-dependent protein refolding / Signaling by ERBB2 / telomere maintenance via telomerase / HSF1-dependent transactivation / positive regulation of cell size / response to unfolded protein / protein unfolding / negative regulation of extrinsic apoptotic signaling pathway in absence of ligand / chaperone-mediated protein complex assembly / Regulation of HSF1-mediated heat shock response / HSF1 activation / regulation of protein-containing complex assembly / Attenuation phase / cellular response to unfolded protein / RHOBTB2 GTPase cycle / estrogen response element binding / FOXO-mediated transcription of oxidative stress, metabolic and neuronal genes / cellular response to dexamethasone stimulus / eNOS activation / axonal growth cone / positive regulation of lamellipodium assembly / DNA polymerase binding / nuclear receptor-mediated steroid hormone signaling pathway / core promoter sequence-specific DNA binding / ATP metabolic process / Tetrahydrobiopterin (BH4) synthesis, recycling, salvage and regulation / inclusion body / positive regulation of defense response to virus by host / Loss of Nlp from mitotic centrosomes / Loss of proteins required for interphase microtubule organization from the centrosome / Recruitment of mitotic centrosome proteins and complexes 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: 38.0 Å | |||||||||
![]() | Kirschke E / Goswami D / Southworth DR / Griffin PR / Agard DA | |||||||||
![]() | ![]() タイトル: Glucocorticoid receptor function regulated by coordinated action of the Hsp90 and Hsp70 chaperone cycles. 著者: Elaine Kirschke / Devrishi Goswami / Daniel Southworth / Patrick R Griffin / David A Agard / ![]() 要旨: The glucocorticoid receptor (GR), like many signaling proteins, depends on the Hsp90 molecular chaperone for in vivo function. Although Hsp90 is required for ligand binding in vivo, purified apo GR ...The glucocorticoid receptor (GR), like many signaling proteins, depends on the Hsp90 molecular chaperone for in vivo function. Although Hsp90 is required for ligand binding in vivo, purified apo GR is capable of binding ligand with no enhancement from Hsp90. We reveal that Hsp70, known to facilitate client delivery to Hsp90, inactivates GR through partial unfolding, whereas Hsp90 reverses this inactivation. Full recovery of ligand binding requires ATP hydrolysis on Hsp90 and the Hop and p23 cochaperones. Surprisingly, Hsp90 ATP hydrolysis appears to regulate client transfer from Hsp70, likely through a coupling of the two chaperone's ATP cycles. Such coupling is embodied in contacts between Hsp90 and Hsp70 in the GR:Hsp70:Hsp90:Hop complex imaged by cryoelectron microscopy. Whereas GR released from Hsp70 is aggregation prone, release from Hsp90 protects GR from aggregation and enhances its ligand affinity. Together, this illustrates how coordinated chaperone interactions can enhance stability, function, and regulation. | |||||||||
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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マップデータ | ![]() | 3.7 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 13.9 KB 13.9 KB | 表示 表示 | ![]() |
画像 | ![]() | 73.2 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
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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 | Reconstruction of client-bound Hsp90-Hsp70-Hop chaperone complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 2.4 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
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試料の構成要素
-全体 : Human chaperones Hsp90 and Hsp70 bound with the human co-chaperon...
全体 | 名称: Human chaperones Hsp90 and Hsp70 bound with the human co-chaperone Hop and the ligand binding domain of the human glucocorticoid receptor N-terminally fused with the maltose binding protein |
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要素 |
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-超分子 #1000: Human chaperones Hsp90 and Hsp70 bound with the human co-chaperon...
超分子 | 名称: Human chaperones Hsp90 and Hsp70 bound with the human co-chaperone Hop and the ligand binding domain of the human glucocorticoid receptor N-terminally fused with the maltose binding protein タイプ: sample / ID: 1000 詳細: Complex was purified using size exclusion chromatography. 集合状態: One homodimer of Hsp90 bound to one Hsp70, one Hop, and one MBP-GRLBD Number unique components: 4 |
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分子量 | 理論値: 380 KDa |
-分子 #1: Heat shock protein 90-alpha
分子 | 名称: Heat shock protein 90-alpha / タイプ: protein_or_peptide / ID: 1 / Name.synonym: Hsp90 / コピー数: 1 / 集合状態: Dimer / 組換発現: Yes |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 84.66 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | UniProtKB: Heat shock protein HSP 90-alpha / InterPro: Heat shock protein Hsp90 family |
-分子 #2: Heat shock 70 kDa protein
分子 | 名称: Heat shock 70 kDa protein / タイプ: protein_or_peptide / ID: 2 / Name.synonym: Hsp70 / コピー数: 1 / 集合状態: monomer / 組換発現: Yes |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 70.052 KDa |
組換発現 | 生物種: ![]() ![]() 組換細胞: Sf9 |
配列 | UniProtKB: Heat shock 70 kDa protein 1B / InterPro: Heat shock protein 70 family |
-分子 #3: Glucocorticoid Receptor Ligand Binding Domain
分子 | 名称: Glucocorticoid Receptor Ligand Binding Domain / タイプ: protein_or_peptide / ID: 3 / Name.synonym: GRLBD 詳細: Maltose Binding Protein (MBP) tag attached to N-terminus コピー数: 1 / 集合状態: monomer / 組換発現: Yes |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 74.926 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | UniProtKB: Glucocorticoid receptor / InterPro: Glucocorticoid receptor |
-分子 #4: Stress-induced-phosphoprotein 1
分子 | 名称: Stress-induced-phosphoprotein 1 / タイプ: protein_or_peptide / ID: 4 / Name.synonym: Hop / コピー数: 1 / 集合状態: monomer / 組換発現: Yes |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 62.639 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | UniProtKB: Stress-induced-phosphoprotein 1 / InterPro: Heat shock chaperonin-binding |
-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
緩衝液 | pH: 7.5 詳細: 50 mM HEPES, 50 mM KCl, 2 mM DTT, 5 mM MgCl2, 0.2 mM ADP |
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グリッド | 詳細: C-flat holey carbon grid, 2 micron hole, 0.5 micron space |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 70 % / チャンバー内温度: 75 K / 装置: FEI VITROBOT MARK I / 手法: two one-second blots before plunging |
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電子顕微鏡法
顕微鏡 | FEI TECNAI F20 |
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アライメント法 | Legacy - 非点収差: Astigmatism was corrected at 250,000 times magnification. |
日付 | 2011年9月21日 |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: TVIPS TEMCAM-F816 (8k x 8k) 実像数: 214 / 平均電子線量: 30 e/Å2 |
電子線 | 加速電圧: 120 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.2 mm / 最大 デフォーカス(公称値): 4.0 µm / 最小 デフォーカス(公称値): 2.2 µm / 倍率(公称値): 60000 |
試料ステージ | 試料ホルダーモデル: OTHER |
実験機器 | ![]() モデル: Tecnai F20 / 画像提供: FEI Company |
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画像解析
詳細 | The particles were manually selected. |
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CTF補正 | 詳細: CTF Find |
最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 38.0 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: EMAN / 使用した粒子像数: 10149 |
最終 角度割当 | 詳細: EMAN: 15 |