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基本情報
登録情報 | データベース: PDB / ID: 8bw9 | ||||||||||||
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タイトル | Cryo-EM structure of the RAF activating complex KSR-MEK-CNK-HYP | ||||||||||||
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![]() | SIGNALING PROTEIN / Kinase suppressor of Ras (KSR) / Dual specificity mitogen-activated protein kinase kinase (MEK) / Connector enhancer of KSR (CNK) / Protein Aveugle (AVE) / Hyphen protein (HYP) / protein complex | ||||||||||||
機能・相同性 | ![]() hemocyte differentiation / MAPK3 (ERK1) activation / MAPK1 (ERK2) activation / Frs2-mediated activation / Signal transduction by L1 / Negative feedback regulation of MAPK pathway / terminal region determination / RAF activation / Phosphorylation of CI / compound eye cone cell differentiation ...hemocyte differentiation / MAPK3 (ERK1) activation / MAPK1 (ERK2) activation / Frs2-mediated activation / Signal transduction by L1 / Negative feedback regulation of MAPK pathway / terminal region determination / RAF activation / Phosphorylation of CI / compound eye cone cell differentiation / Phosphorylation of SMO / terminal branching, open tracheal system / torso signaling pathway / photoreceptor cell development / tracheal outgrowth, open tracheal system / R7 cell fate commitment / compound eye photoreceptor cell differentiation / sevenless signaling pathway / Phosphorylation of PER and TIM / MAP2K and MAPK activation / eye photoreceptor cell differentiation / epithelial cell migration, open tracheal system / imaginal disc-derived wing vein specification / border follicle cell migration / imaginal disc-derived wing morphogenesis / anterior/posterior axis specification, embryo / cellular response to X-ray / mitogen-activated protein kinase kinase / MAP-kinase scaffold activity / mitogen-activated protein kinase kinase kinase binding / mitotic DNA replication checkpoint signaling / dorsal/ventral pattern formation / positive regulation of Ras protein signal transduction / mitotic G2 DNA damage checkpoint signaling / MAP kinase kinase activity / fibroblast growth factor receptor signaling pathway / vascular endothelial growth factor receptor signaling pathway / enzyme regulator activity / condensed chromosome / cell surface receptor protein tyrosine kinase signaling pathway / visual perception / ERK1 and ERK2 cascade / determination of adult lifespan / epidermal growth factor receptor signaling pathway / cytoplasmic side of plasma membrane / receptor tyrosine kinase binding / kinase binding / MAPK cascade / insulin receptor signaling pathway / apical part of cell / cell cortex / scaffold protein binding / protein tyrosine kinase activity / defense response to virus / Ras protein signal transduction / positive regulation of ERK1 and ERK2 cascade / non-specific serine/threonine protein kinase / protein kinase activity / protein phosphorylation / protein serine kinase activity / protein serine/threonine kinase activity / ATP binding / metal ion binding / plasma membrane / cytoplasm / cytosol 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: 3.32 Å | ||||||||||||
![]() | Maisonneuve, P. / Fronzes, R. / Sicheri, F. | ||||||||||||
資金援助 | ![]() ![]()
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![]() | ![]() タイトル: The CNK-HYP scaffolding complex promotes RAF activation by enhancing KSR-MEK interaction. 著者: Pierre Maisonneuve / Malha Sahmi / Fanny Bergeron-Labrecque / Xianjie Iris Ma / Juliette Queguiner / Geneviève Arseneault / Martin Lefrançois / Igor Kurinov / Rémi Fronzes / Frank Sicheri / Marc Therrien / ![]() ![]() ![]() 要旨: The RAS-MAPK pathway regulates cell proliferation, differentiation and survival, and its dysregulation is associated with cancer development. The pathway minimally comprises the small GTPase RAS and ...The RAS-MAPK pathway regulates cell proliferation, differentiation and survival, and its dysregulation is associated with cancer development. The pathway minimally comprises the small GTPase RAS and the kinases RAF, MEK and ERK. Activation of RAF by RAS is notoriously intricate and remains only partially understood. There are three RAF isoforms in mammals (ARAF, BRAF and CRAF) and two related pseudokinases (KSR1 and KSR2). RAS-mediated activation of RAF depends on an allosteric mechanism driven by the dimerization of its kinase domain. Recent work on human RAFs showed that MEK binding to KSR1 promotes KSR1-BRAF heterodimerization, which leads to the phosphorylation of free MEK molecules by BRAF. Similar findings were made with the single Drosophila RAF homolog. Here we show that the fly scaffold proteins CNK and HYP stabilize the KSR-MEK interaction, which in turn enhances RAF-KSR heterodimerization and RAF activation. The cryogenic electron microscopy structure of the minimal KSR-MEK-CNK-HYP complex reveals a ring-like arrangement of the CNK-HYP complex allowing CNK to simultaneously engage KSR and MEK, thus stabilizing the binary interaction. Together, these results illuminate how CNK contributes to RAF activation by stimulating the allosteric function of KSR and highlight the diversity of mechanisms impacting RAF dimerization as well as the regulatory potential of the KSR-MEK interaction. | ||||||||||||
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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PDBx/mmCIF形式 | ![]() | 223.3 KB | 表示 | ![]() |
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PDB形式 | ![]() | 145.8 KB | 表示 | ![]() |
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-検証レポート
文書・要旨 | ![]() | 1.6 MB | 表示 | ![]() |
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文書・詳細版 | ![]() | 1.6 MB | 表示 | |
XML形式データ | ![]() | 38.1 KB | 表示 | |
CIF形式データ | ![]() | 53.8 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 | ![]() 16281MC ![]() 8bw8C 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
-タンパク質 , 4種, 4分子 ABCD
#1: タンパク質 | 分子量: 13143.911 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) ![]() ![]() 遺伝子: ave, CG30476 / 発現宿主: ![]() ![]() |
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#2: タンパク質 | 分子量: 38534.203 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) ![]() ![]() 遺伝子: cnk, anon-WO0140519.129, anon-WO0257455.27, CNK, Cnk, Dmel\CG6556, EC2-3, EK2-3, l(2)k16314, sag, CG6556, Dmel_CG6556 発現宿主: ![]() ![]() |
#3: タンパク質 | 分子量: 43926.496 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) ![]() ![]() 遺伝子: Dsor1, CG15793 発現宿主: ![]() ![]() 参照: UniProt: Q24324, mitogen-activated protein kinase kinase |
#4: タンパク質 | 分子量: 36474.898 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) ![]() ![]() 遺伝子: ksr, CG2899 発現宿主: ![]() ![]() 参照: UniProt: Q24170 |
-非ポリマー , 3種, 3分子 ![](data/chem/img/ANP.gif)
![](data/chem/img/QOM.gif)
![](data/chem/img/MG.gif)
![](data/chem/img/QOM.gif)
![](data/chem/img/MG.gif)
#5: 化合物 | ChemComp-ANP / |
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#6: 化合物 | ChemComp-QOM / |
#7: 化合物 | ChemComp-MG / |
-詳細
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: Quaternary complex of the kinase domains of KSR and MEK bound to the scaffolding complex CNK-HYP タイプ: COMPLEX / 詳細: Mg2+/ANP and Trametinib ligands bound to MEK / Entity ID: #1-#4 / 由来: RECOMBINANT | |||||||||||||||||||||||||
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分子量 | 値: 0.132 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 / 詳細: AMPPNP 25mM Trametinib 0.05mM | |||||||||||||||||||||||||
緩衝液成分 |
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試料 | 濃度: 0.79 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: COPPER / グリッドのサイズ: 200 divisions/in. / グリッドのタイプ: Quantifoil 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: 277.15 K / 詳細: 3.5 sec blot time 1 sec drain time |
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電子顕微鏡撮影
実験機器 | ![]() モデル: Talos Arctica / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TALOS ARCTICA |
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): 26398 nm / 最小 デフォーカス(公称値): 5212 nm |
撮影 | 電子線照射量: 1.15 e/Å2 / 検出モード: COUNTING フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 実像数: 44 |
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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 | ||||||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||||||
3次元再構成 | 解像度: 3.32 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 141531 / 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: 119.78 Å2 | ||||||||||||||||||||||||
拘束条件 |
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