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基本情報
登録情報 | データベース: PDB / ID: 7z37 | |||||||||||||||
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タイトル | Structure of the RAF1-HSP90-CDC37 complex (RHC-II) | |||||||||||||||
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![]() | TRANSFERASE / RAF1-HSP90-CDC37 / complex / proto-oncogene / serine/threonine kinase / cancer / chaperone | |||||||||||||||
機能・相同性 | ![]() regulation of type II interferon-mediated signaling pathway / : / HSP90-CDC37 chaperone complex / positive regulation of cyclin-dependent protein kinase activity / death-inducing signaling complex assembly / positive regulation of mitophagy in response to mitochondrial depolarization / negative regulation of proteasomal protein catabolic process / Aryl hydrocarbon receptor signalling / aryl hydrocarbon receptor complex / dynein axonemal particle ...regulation of type II interferon-mediated signaling pathway / : / HSP90-CDC37 chaperone complex / positive regulation of cyclin-dependent protein kinase activity / death-inducing signaling complex assembly / positive regulation of mitophagy in response to mitochondrial depolarization / negative regulation of proteasomal protein catabolic process / Aryl hydrocarbon receptor signalling / aryl hydrocarbon receptor complex / dynein axonemal particle / intermediate filament cytoskeleton organization / histone methyltransferase binding / type B pancreatic cell proliferation / regulation of Rho protein signal transduction / SHOC2 M1731 mutant abolishes MRAS complex function / Gain-of-function MRAS complexes activate RAF signaling / positive regulation of protein localization to cell surface / Rap1 signalling / ATP-dependent protein binding / protein kinase regulator activity / regulation of cell motility / protein folding chaperone complex / insulin secretion involved in cellular response to glucose stimulus / negative regulation of protein metabolic process / positive regulation of tau-protein kinase activity / Negative feedback regulation of MAPK pathway / post-transcriptional regulation of gene expression / telomerase holoenzyme complex assembly / Respiratory syncytial virus genome replication / regulation of cyclin-dependent protein serine/threonine kinase activity / Uptake and function of diphtheria toxin / GP1b-IX-V activation signalling / IFNG signaling activates MAPKs / 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 / TPR domain binding / ERBB2-ERBB3 signaling pathway / positive regulation of transforming growth factor beta receptor signaling pathway / regulation of cell differentiation / Assembly and release of respiratory syncytial virus (RSV) virions / face development / pseudopodium / dendritic growth cone / somatic stem cell population maintenance / regulation of type I interferon-mediated signaling pathway / positive regulation of phosphoprotein phosphatase activity / thyroid gland development / neurotrophin TRK receptor signaling pathway / Sema3A PAK dependent Axon repulsion / The NLRP3 inflammasome / regulation of protein ubiquitination / HSF1-dependent transactivation / response to unfolded protein / negative regulation of proteasomal ubiquitin-dependent protein catabolic process / telomere maintenance via telomerase / extrinsic apoptotic signaling pathway via death domain receptors / chaperone-mediated protein complex assembly / MAP kinase kinase kinase activity / HSF1 activation / Attenuation phase / protein targeting / cellular response to interleukin-4 / negative regulation of protein-containing complex assembly / RHOBTB2 GTPase cycle / Purinergic signaling in leishmaniasis infection / Schwann cell development / axonal growth cone / type II interferon-mediated signaling pathway / DNA polymerase binding / supramolecular fiber organization / negative regulation of extrinsic apoptotic signaling pathway via death domain receptors / Signaling by ERBB2 / : / heat shock protein binding / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / activation of adenylate cyclase activity / response to muscle stretch / nitric-oxide synthase regulator activity / myelination / CD209 (DC-SIGN) signaling / Constitutive Signaling by Overexpressed ERBB2 / ESR-mediated signaling / insulin-like growth factor receptor signaling pathway / thymus development / positive regulation of cell differentiation / ATP-dependent protein folding chaperone / peptide binding / Signaling by ERBB2 TMD/JMD mutants / RAF activation / wound healing / Hsp90 protein binding / Signaling by high-kinase activity BRAF mutants 類似検索 - 分子機能 | |||||||||||||||
生物種 | ![]() | |||||||||||||||
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: 3.67 Å | |||||||||||||||
![]() | Mesa, P. / Garcia-Alonso, S. / Barbacid, M. / Montoya, G. | |||||||||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Structure of the RAF1-HSP90-CDC37 complex reveals the basis of RAF1 regulation. 著者: Sara García-Alonso / Pablo Mesa / Laura de la Puente Ovejero / Gonzalo Aizpurua / Carmen G Lechuga / Eduardo Zarzuela / Clara M Santiveri / Manuel Sanclemente / Javier Muñoz / Mónica ...著者: Sara García-Alonso / Pablo Mesa / Laura de la Puente Ovejero / Gonzalo Aizpurua / Carmen G Lechuga / Eduardo Zarzuela / Clara M Santiveri / Manuel Sanclemente / Javier Muñoz / Mónica Musteanu / Ramón Campos-Olivas / Jorge Martínez-Torrecuadrada / Mariano Barbacid / Guillermo Montoya / ![]() ![]() 要旨: RAF kinases are RAS-activated enzymes that initiate signaling through the MAPK cascade to control cellular proliferation, differentiation, and survival. Here, we describe the structure of the full- ...RAF kinases are RAS-activated enzymes that initiate signaling through the MAPK cascade to control cellular proliferation, differentiation, and survival. Here, we describe the structure of the full-length RAF1 protein in complex with HSP90 and CDC37 obtained by cryoelectron microscopy. The reconstruction reveals a RAF1 kinase with an unfolded N-lobe separated from its C-lobe. The hydrophobic core of the N-lobe is trapped in the HSP90 dimer, while CDC37 wraps around the chaperone and interacts with the N- and C-lobes of the kinase. The structure indicates how CDC37 can discriminate between the different members of the RAF family. Our structural analysis also reveals that the folded RAF1 assembles with 14-3-3 dimers, suggesting that after folding RAF1 follows a similar activation as B-RAF. Finally, disruption of the interaction between CDC37 and the DFG segment of RAF1 unveils potential vulnerabilities in attempting the pharmacological degradation of RAF1 for therapeutic purposes. | |||||||||||||||
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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-検証レポート
文書・要旨 | ![]() | 1.1 MB | 表示 | ![]() |
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文書・詳細版 | ![]() | 1.1 MB | 表示 | |
XML形式データ | ![]() | 63.2 KB | 表示 | |
CIF形式データ | ![]() | 88 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 | ![]() 14472MC ![]() 7z38C M: このデータのモデリングに利用したマップデータ C: 同じ文献を引用 ( |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
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リンク
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集合体
登録構造単位 | ![]()
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要素
#1: タンパク質 | 分子量: 84371.281 Da / 分子数: 2 / 由来タイプ: 組換発現 / 詳細: N-terminal HA-tag + HSP90-beta / 由来: (組換発現) ![]() ![]() #2: タンパク質 | | 分子量: 74409.953 Da / 分子数: 1 / 由来タイプ: 組換発現 / 詳細: RAF1 + linker Gly-Ser-Ala + C-terminal StrepTagII / 由来: (組換発現) ![]() ![]() 参照: UniProt: P04049, non-specific serine/threonine protein kinase #3: タンパク質 | | 分子量: 46535.516 Da / 分子数: 1 / 由来タイプ: 組換発現 / 詳細: CDC37 + C-terminal Myc-DDK tag / 由来: (組換発現) ![]() ![]() #4: 化合物 | 研究の焦点であるリガンドがあるか | Y | |
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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 |
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試料調製
構成要素 | 名称: RAF1-HSP90-CDC37 complex, RHC-II / タイプ: COMPLEX / Entity ID: #1-#3 / 由来: RECOMBINANT | ||||||||||||||||||||||||||||||
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分子量 | 値: 0.28913380 MDa / 実験値: NO | ||||||||||||||||||||||||||||||
由来(天然) | 生物種: ![]() | ||||||||||||||||||||||||||||||
由来(組換発現) | 生物種: ![]() | ||||||||||||||||||||||||||||||
緩衝液 | pH: 7.5 | ||||||||||||||||||||||||||||||
緩衝液成分 |
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試料 | 濃度: 0.3 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | ||||||||||||||||||||||||||||||
試料支持 | 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: UltrAuFoil R2/2 | ||||||||||||||||||||||||||||||
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: 277 K / 詳細: blot for 3 seconds before plunging |
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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: ![]() |
電子レンズ | モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2500 nm / 最小 デフォーカス(公称値): 500 nm / Cs: 2.7 mm / C2レンズ絞り径: 100 µm |
試料ホルダ | 凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
撮影 | 電子線照射量: 30 e/Å2 / 検出モード: COUNTING フィルム・検出器のモデル: FEI FALCON III (4k x 4k) 撮影したグリッド数: 1 / 実像数: 8138 詳細: 8138 images (4430 non-tilted and 3708 30 degrees tilted) |
画像スキャン | 横: 4096 / 縦: 4096 |
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解析
ソフトウェア |
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EMソフトウェア |
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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: 3760000 詳細: 3760k in total: 1353k RAF1:HSP90:CDC37 complex, 1127k 14-3-3 dimer, RAF1:14-3-3 complex 1245k | |||||||||||||||||||||||||||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | |||||||||||||||||||||||||||||||||||||||||||||
3次元再構成 | 解像度: 3.67 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 46000 / 対称性のタイプ: POINT | |||||||||||||||||||||||||||||||||||||||||||||
精密化 | 交差検証法: NONE 立体化学のターゲット値: GeoStd + Monomer Library + CDL v1.2 | |||||||||||||||||||||||||||||||||||||||||||||
原子変位パラメータ | Biso mean: 110.87 Å2 | |||||||||||||||||||||||||||||||||||||||||||||
拘束条件 |
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