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基本情報
登録情報 | データベース: PDB / ID: 3jbq | ||||||
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タイトル | Domain Organization and Conformational Plasticity of the G Protein Effector, PDE6 | ||||||
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![]() | HYDROLASE/IMMUNE SYSTEM / Phosphodiesterase / photoreceptor / PDE6 / HYDROLASE-IMMUNE SYSTEM complex | ||||||
機能・相同性 | ![]() cGMP effects / Smooth Muscle Contraction / RHOBTB1 GTPase cycle / cyclic-nucleotide phosphodiesterase activity / GMP catabolic process / cellular response to macrophage colony-stimulating factor stimulus / 3',5'-cyclic-GMP phosphodiesterase / cellular response to cGMP / positive regulation of G protein-coupled receptor signaling pathway / Inactivation, recovery and regulation of the phototransduction cascade ...cGMP effects / Smooth Muscle Contraction / RHOBTB1 GTPase cycle / cyclic-nucleotide phosphodiesterase activity / GMP catabolic process / cellular response to macrophage colony-stimulating factor stimulus / 3',5'-cyclic-GMP phosphodiesterase / cellular response to cGMP / positive regulation of G protein-coupled receptor signaling pathway / Inactivation, recovery and regulation of the phototransduction cascade / Activation of the phototransduction cascade / positive regulation of vascular permeability / ion binding / cellular response to granulocyte macrophage colony-stimulating factor stimulus / response to stimulus / negative regulation of vascular permeability / establishment of endothelial barrier / negative regulation of cAMP-mediated signaling / regulation of mitochondrion organization / Ca2+ pathway / positive regulation of epidermal growth factor receptor signaling pathway / photoreceptor outer segment membrane / cGMP-stimulated cyclic-nucleotide phosphodiesterase activity / 3',5'-cyclic-nucleotide phosphodiesterase / negative regulation of cGMP-mediated signaling / cGMP-mediated signaling / cGMP catabolic process / cGMP binding / 3',5'-cyclic-GMP phosphodiesterase activity / 3',5'-cyclic-AMP phosphodiesterase activity / regulation of cAMP-mediated signaling / visual perception / cAMP-mediated signaling / synaptic membrane / photoreceptor disc membrane / positive regulation of inflammatory response / cellular response to mechanical stimulus / presynaptic membrane / mitochondrial outer membrane / mitochondrial inner membrane / molecular adaptor activity / mitochondrial matrix / positive regulation of gene expression / perinuclear region of cytoplasm / Golgi apparatus / negative regulation of transcription by RNA polymerase II / endoplasmic reticulum / signal transduction / protein homodimerization activity / zinc ion binding / nucleus / 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: 11 Å | ||||||
![]() | Zhang, Z. / He, F. / Constantine, R. / Baker, M.L. / Baehr, W. / Schmid, M.F. / Wensel, T.G. / Agosto, M.A. | ||||||
![]() | ![]() タイトル: Domain organization and conformational plasticity of the G protein effector, PDE6. 著者: Zhixian Zhang / Feng He / Ryan Constantine / Matthew L Baker / Wolfgang Baehr / Michael F Schmid / Theodore G Wensel / Melina A Agosto / ![]() 要旨: The cGMP phosphodiesterase of rod photoreceptor cells, PDE6, is the key effector enzyme in phototransduction. Two large catalytic subunits, PDE6α and -β, each contain one catalytic domain and two ...The cGMP phosphodiesterase of rod photoreceptor cells, PDE6, is the key effector enzyme in phototransduction. Two large catalytic subunits, PDE6α and -β, each contain one catalytic domain and two non-catalytic GAF domains, whereas two small inhibitory PDE6γ subunits allow tight regulation by the G protein transducin. The structure of holo-PDE6 in complex with the ROS-1 antibody Fab fragment was determined by cryo-electron microscopy. The ∼11 Å map revealed previously unseen features of PDE6, and each domain was readily fit with high resolution structures. A structure of PDE6 in complex with prenyl-binding protein (PrBP/δ) indicated the location of the PDE6 C-terminal prenylations. Reconstructions of complexes with Fab fragments bound to N or C termini of PDE6γ revealed that PDE6γ stretches from the catalytic domain at one end of the holoenzyme to the GAF-A domain at the other. Removal of PDE6γ caused dramatic structural rearrangements, which were reversed upon its restoration. | ||||||
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構造の表示
ムービー |
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構造ビューア | 分子: ![]() ![]() |
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-検証レポート
文書・要旨 | ![]() | 666.4 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 671.4 KB | 表示 | |
XML形式データ | ![]() | 54.2 KB | 表示 | |
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-関連構造データ
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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
-Phosphodiesterase ... , 2種, 4分子 BFDX
#3: タンパク質 | 分子量: 38282.086 Da / 分子数: 2 / 断片: SEE REMARK 999 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #4: タンパク質・ペプチド | 分子量: 2185.436 Da / 分子数: 2 / 断片: UNP residues 70-87 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() |
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-タンパク質 , 2種, 4分子 CG12
#5: タンパク質 | 分子量: 20671.434 Da / 分子数: 2 / 断片: SEE REMARK 999 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #6: タンパク質 | 分子量: 21554.822 Da / 分子数: 2 / 断片: SEE REMARK 999 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() |
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-抗体 , 2種, 4分子 LlHh
#1: 抗体 | 分子量: 23540.990 Da / 分子数: 2 / 断片: Fab / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #2: 抗体 | 分子量: 23781.471 Da / 分子数: 2 / 断片: Fab / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() |
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-詳細
配列の詳細 | THE IMAGED PDE6 WAS DERIVED FROM BOS TAURUS, BUT SOME MODELED SEQUENCES ARE FROM HOMO SAPIENS ...THE IMAGED PDE6 WAS DERIVED FROM BOS TAURUS, BUT SOME MODELED SEQUENCES ARE FROM HOMO SAPIENS (CHAINS B, C, F, G) OR GALLUS GALLUS (CHAINS 1, 2). THE PHOSPHODIE |
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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 |
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分子量 | 値: 0.32 MDa / 実験値: NO | ||||||||||||||||||||
緩衝液 | 名称: 20 mM sodium phosphate, 150 mM sodium chloride / pH: 7.5 / 詳細: 20 mM sodium phosphate, 150 mM sodium chloride | ||||||||||||||||||||
試料 | 濃度: 0.5 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: 400 mesh glow-discharged Quantifoil grids with 2.0 Angstrom holes | ||||||||||||||||||||
急速凍結 | 装置: FEI VITROBOT MARK III / 凍結剤: ETHANE / Temp: 93 K / 湿度: 95 % 詳細: Applied 3 uL of sample per grid and blotted for 1 second before plunging into liquid ethane (FEI VITROBOT MARK III). 手法: Applied 3 ul of sample per grid, blotted for 1 second before plunging |
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電子顕微鏡撮影
顕微鏡 | モデル: JEOL 2010F / 日付: 2011年8月8日 / 詳細: Parallel beam illumination |
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電子銃 | 電子線源: ![]() |
電子レンズ | モード: BRIGHT FIELD / 倍率(公称値): 60000 X / Cs: 2 mm 非点収差: Objective lens astigmatism was corrected at 100,000 times magnification. カメラ長: 0 mm |
試料ホルダ | 試料ホルダーモデル: GATAN LIQUID NITROGEN / 最高温度: 94 K |
撮影 | 電子線照射量: 15 e/Å2 フィルム・検出器のモデル: GATAN ULTRASCAN 4000 (4k x 4k) |
電子光学装置 | エネルギーフィルター名称: FEI |
放射 | プロトコル: SINGLE WAVELENGTH / 単色(M)・ラウエ(L): M / 散乱光タイプ: x-ray |
放射波長 | 相対比: 1 |
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解析
EMソフトウェア |
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CTF補正 | 詳細: by particle using CtFit | ||||||||||||||||||||||||||||
対称性 | 点対称性: C2 (2回回転対称) | ||||||||||||||||||||||||||||
3次元再構成 | 手法: projection matching / 解像度: 11 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 12373 / ピクセルサイズ(公称値): 1.81 Å / ピクセルサイズ(実測値): 1.81 Å 詳細: A total of 21100 particles were picked from ice images and CTF-corrected using Ctfit. Initial three-dimensional models were generated from either reference-free class averages or from a ...詳細: A total of 21100 particles were picked from ice images and CTF-corrected using Ctfit. Initial three-dimensional models were generated from either reference-free class averages or from a cylinder. The two models were essentially similar. Three noise-seeded models were generated and used as initial models in Multirefine. A model with two Ros-1 Fabs bound emerged from a population of 15000 particles and was subjected to further refinement using standard iterative projection matching, class averaging, and Fourier reconstruction. The final three-dimensional map with C2 symmetry was generated from 12373 particles. クラス平均像の数: 20 / 対称性のタイプ: POINT | ||||||||||||||||||||||||||||
原子モデル構築 | プロトコル: RIGID BODY FIT / 空間: REAL 詳細: REFINEMENT PROTOCOL--rigid body refinement DETAILS--The domains were fit using Chimera. The model was refined using Phenix real-space refinement. | ||||||||||||||||||||||||||||
原子モデル構築 |
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精密化ステップ | サイクル: LAST
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