+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 8ttb | ||||||||||||
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タイトル | Cryo-EM structure of the PP2A:B55-ARPP19 complex | ||||||||||||
要素 |
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キーワード | SIGNALING PROTEIN / HYDROLASE (加水分解酵素) / Protein Phosphatase 2A:B55 holoenzyme / ARPP19 inhibitor / cell cycle regulation (細胞周期) | ||||||||||||
機能・相同性 | 機能・相同性情報 phosphatase inhibitor activity / negative regulation of protein dephosphorylation / meiotic spindle elongation / Integration of energy metabolism / PP2A-mediated dephosphorylation of key metabolic factors / regulation of microtubule binding / MASTL Facilitates Mitotic Progression / mitotic sister chromatid separation / regulation of meiotic cell cycle process involved in oocyte maturation / protein phosphatase type 2A complex ...phosphatase inhibitor activity / negative regulation of protein dephosphorylation / meiotic spindle elongation / Integration of energy metabolism / PP2A-mediated dephosphorylation of key metabolic factors / regulation of microtubule binding / MASTL Facilitates Mitotic Progression / mitotic sister chromatid separation / regulation of meiotic cell cycle process involved in oocyte maturation / protein phosphatase type 2A complex / meiotic sister chromatid cohesion, centromeric / peptidyl-serine dephosphorylation / : / peptidyl-threonine dephosphorylation / negative regulation of tyrosine phosphorylation of STAT protein / positive regulation of microtubule binding / Regulation of glycolysis by fructose 2,6-bisphosphate metabolism / Inhibition of replication initiation of damaged DNA by RB1/E2F1 / female meiotic nuclear division / protein antigen binding / ceramide metabolic process / protein phosphatase regulator activity / GABA receptor binding / negative regulation of epithelial to mesenchymal transition / APC truncation mutants have impaired AXIN binding / AXIN missense mutants destabilize the destruction complex / Truncations of AMER1 destabilize the destruction complex / Initiation of Nuclear Envelope (NE) Reformation / ERKs are inactivated / response to morphine / positive regulation of extrinsic apoptotic signaling pathway in absence of ligand / Beta-catenin phosphorylation cascade / Signaling by GSK3beta mutants / CTNNB1 S33 mutants aren't phosphorylated / CTNNB1 S37 mutants aren't phosphorylated / CTNNB1 S45 mutants aren't phosphorylated / CTNNB1 T41 mutants aren't phosphorylated / regulation of Wnt signaling pathway / regulation of growth / Disassembly of the destruction complex and recruitment of AXIN to the membrane / protein phosphatase inhibitor activity / negative regulation of glycolytic process through fructose-6-phosphate / positive regulation of NLRP3 inflammasome complex assembly / myosin phosphatase activity / protein serine/threonine phosphatase activity / CTLA4 inhibitory signaling / Platelet sensitization by LDL / negative regulation of MAPK cascade / protein-serine/threonine phosphatase / regulation of cell differentiation / T cell homeostasis / ERK/MAPK targets / regulation of G1/S transition of mitotic cell cycle / phosphoprotein phosphatase activity / regulation of DNA replication / mesoderm development / chromosome, centromeric region / DARPP-32 events / lateral plasma membrane / negative regulation of phosphatidylinositol 3-kinase/protein kinase B signal transduction / potassium channel regulator activity / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / regulation of cell adhesion / Cyclin A/B1/B2 associated events during G2/M transition / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / positive regulation of gluconeogenesis / Loss of Nlp from mitotic centrosomes / Loss of proteins required for interphase microtubule organization from the centrosome / Recruitment of mitotic centrosome proteins and complexes / Resolution of Sister Chromatid Cohesion / Recruitment of NuMA to mitotic centrosomes / Anchoring of the basal body to the plasma membrane / AURKA Activation by TPX2 / protein dephosphorylation / RNA splicing / meiotic cell cycle / protein phosphatase 2A binding / response to organic substance / protein tyrosine phosphatase activity / chromosome segregation / RHO GTPases Activate Formins / positive regulation of glucose import / response to lead ion / regulation of protein phosphorylation / Spry regulation of FGF signaling / RAF activation / Degradation of beta-catenin by the destruction complex / PKR-mediated signaling / tau protein binding / positive regulation of protein serine/threonine kinase activity / negative regulation of cell growth / 紡錘体 / Negative regulation of MAPK pathway / Separation of Sister Chromatids / Cyclin D associated events in G1 / microtubule cytoskeleton / G2/M transition of mitotic cell cycle / Regulation of PLK1 Activity at G2/M Transition 類似検索 - 分子機能 | ||||||||||||
生物種 | Homo sapiens (ヒト) | ||||||||||||
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: 2.77 Å | ||||||||||||
データ登録者 | Fuller, J.R. / Padi, S.K.R. / Peti, W. / Page, R. | ||||||||||||
資金援助 | 米国, 3件
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引用 | ジャーナル: Nature / 年: 2024 タイトル: Cryo-EM structures of PP2A:B55-FAM122A and PP2A:B55-ARPP19. 著者: Sathish K R Padi / Margaret R Vos / Rachel J Godek / James R Fuller / Thomas Kruse / Jamin B Hein / Jakob Nilsson / Matthew S Kelker / Rebecca Page / Wolfgang Peti / 要旨: Progression through the cell cycle is controlled by regulated and abrupt changes in phosphorylation. Mitotic entry is initiated by increased phosphorylation of mitotic proteins, a process driven by ...Progression through the cell cycle is controlled by regulated and abrupt changes in phosphorylation. Mitotic entry is initiated by increased phosphorylation of mitotic proteins, a process driven by kinases, whereas mitotic exit is achieved by counteracting dephosphorylation, a process driven by phosphatases, especially PP2A:B55. Although the role of kinases in mitotic entry is well established, recent data have shown that mitosis is only successfully initiated when the counterbalancing phosphatases are also inhibited. Inhibition of PP2A:B55 is achieved by the intrinsically disordered proteins ARPP19 and FAM122A. Despite their critical roles in mitosis, the mechanisms by which they achieve PP2A:B55 inhibition is unknown. Here, we report the single-particle cryo-electron microscopy structures of PP2A:B55 bound to phosphorylated ARPP19 and FAM122A. Consistent with our complementary NMR spectroscopy studies, both intrinsically disordered proteins bind PP2A:B55, but do so in highly distinct manners, leveraging multiple distinct binding sites on B55. Our extensive structural, biophysical and biochemical data explain how substrates and inhibitors are recruited to PP2A:B55 and provide a molecular roadmap for the development of therapeutic interventions for PP2A:B55-related diseases. | ||||||||||||
履歴 |
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-構造の表示
構造ビューア | 分子: MolmilJmol/JSmol |
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-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 8ttb.cif.gz | 653.8 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb8ttb.ent.gz | 432.3 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 8ttb.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/tt/8ttb ftp://data.pdbj.org/pub/pdb/validation_reports/tt/8ttb | 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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1 |
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-要素
-Serine/threonine-protein phosphatase 2A ... , 3種, 3分子 ABC
#1: タンパク質 | 分子量: 64957.980 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: PPP2R1A / 発現宿主: Escherichia coli (大腸菌) / 参照: UniProt: P30153 |
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#2: タンパク質 | 分子量: 52044.289 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: PPP2R2A / 細胞株 (発現宿主): expi293 / 発現宿主: Homo sapiens (ヒト) / 参照: UniProt: P63151 |
#3: タンパク質 | 分子量: 35845.375 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: PPP2CA / 細胞株 (発現宿主): expi293 / 発現宿主: Homo sapiens (ヒト) 参照: UniProt: P67775, protein-serine/threonine phosphatase |
-タンパク質 , 1種, 1分子 D
#4: タンパク質 | 分子量: 12620.241 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: ARPP19 / 発現宿主: Escherichia coli (大腸菌) / 参照: UniProt: P56211 |
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-非ポリマー , 2種, 2分子
#5: 化合物 | ChemComp-FE / |
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#6: 化合物 | ChemComp-ZN / |
-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 |
-試料調製
構成要素 | 名称: Quadruple complex of PP2A:B55 (PP2Aa:PP2Ac:B55) bound to thiophosphorylated ARPP19 タイプ: COMPLEX / Entity ID: #1-#4 / 由来: MULTIPLE SOURCES | ||||||||||||||||||||||||||||||
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分子量 | 値: 0.165 MDa / 実験値: NO | ||||||||||||||||||||||||||||||
緩衝液 | pH: 8 詳細: CHAPSO was added only immediately prior to vitrification | ||||||||||||||||||||||||||||||
緩衝液成分 |
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試料 | 濃度: 1.2 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | ||||||||||||||||||||||||||||||
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: 291 K |
-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: FLOOD BEAM |
電子レンズ | モード: BRIGHT FIELDBright-field microscopy / 倍率(公称値): 105000 X / 最大 デフォーカス(公称値): 1900 nm / 最小 デフォーカス(公称値): 700 nm / Calibrated defocus min: 390 nm / 最大 デフォーカス(補正後): 2600 nm / Cs: 2.7 mm / C2レンズ絞り径: 50 µm / アライメント法: ZEMLIN TABLEAU |
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: 70 e/Å2 フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) 撮影したグリッド数: 1 詳細: Camera was operated in CDS mode, with hardware binning of super-resolution pixels, writing movies with 62 frames |
電子光学装置 | エネルギーフィルター名称: GIF Bioquantum / エネルギーフィルタースリット幅: 20 eV |
-解析
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: 1170216 | ||||||||||||||||||||||||||||||||||||||||||||||||
3次元再構成 | 解像度: 2.77 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 52934 / 対称性のタイプ: POINT | ||||||||||||||||||||||||||||||||||||||||||||||||
原子モデル構築 | プロトコル: FLEXIBLE FIT / 空間: REAL / Target criteria: Cross-correlation 詳細: Iterating between manual refinement in Coot and automated real-space refinement in Phenix | ||||||||||||||||||||||||||||||||||||||||||||||||
原子モデル構築 |
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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: 86.62 Å2 | ||||||||||||||||||||||||||||||||||||||||||||||||
拘束条件 |
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