+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 1tvk | ||||||
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タイトル | The binding mode of epothilone A on a,b-tubulin by electron crystallography | ||||||
要素 |
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キーワード | CELL CYCLE / STRUCTURAL PROTEIN / epothilone / taxol / ligand interactions | ||||||
機能・相同性 | 機能・相同性情報 positive regulation of axon guidance / microtubule-based process / 加水分解酵素; 酸無水物に作用; GTPに作用・細胞または細胞小器官の運動に関与 / structural constituent of cytoskeleton / microtubule cytoskeleton organization / microtubule cytoskeleton / nervous system development / mitotic cell cycle / microtubule / hydrolase activity ...positive regulation of axon guidance / microtubule-based process / 加水分解酵素; 酸無水物に作用; GTPに作用・細胞または細胞小器官の運動に関与 / structural constituent of cytoskeleton / microtubule cytoskeleton organization / microtubule cytoskeleton / nervous system development / mitotic cell cycle / microtubule / hydrolase activity / protein heterodimerization activity / GTPase activity / GTP binding / metal ion binding / cytoplasm 類似検索 - 分子機能 | ||||||
生物種 | Bos taurus (ウシ) | ||||||
手法 | 電子線結晶学 / クライオ電子顕微鏡法 / 解像度: 2.89 Å | ||||||
データ登録者 | Nettles, J.H. / Li, H. / Cornett, B. / Krahn, J.M. / Snyder, J.P. / Downing, K.H. | ||||||
引用 | ジャーナル: Science / 年: 2004 タイトル: The binding mode of epothilone A on alpha,beta-tubulin by electron crystallography. 著者: James H Nettles / Huilin Li / Ben Cornett / Joseph M Krahn / James P Snyder / Kenneth H Downing / 要旨: The structure of epothilone A, bound to alpha,beta-tubulin in zinc-stabilized sheets, was determined by a combination of electron crystallography at 2.89 angstrom resolution and nuclear magnetic ...The structure of epothilone A, bound to alpha,beta-tubulin in zinc-stabilized sheets, was determined by a combination of electron crystallography at 2.89 angstrom resolution and nuclear magnetic resonance-based conformational analysis. The complex explains both the broad-based epothilone structure-activity relationship and the known mutational resistance profile. Comparison with Taxol shows that the longstanding expectation of a common pharmacophore is not met, because each ligand exploits the tubulin-binding pocket in a unique and independent manner. | ||||||
履歴 |
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Remark 240 | EXPERIMENT TYPE : ELECTRON DIFFRACTION ELECTRON MICROSCOPE SAMPLE SAMPLE AGGREGATION STATE : TWO- ... EXPERIMENT TYPE : ELECTRON DIFFRACTION ELECTRON MICROSCOPE SAMPLE SAMPLE AGGREGATION STATE : TWO-DIMENSIONAL NAME OF SAMPLE : CRYSTAL TUBULIN- : EPOTHILONE A COMPLEX SAMPLE CONCENTRATION : 2MG/ML SAMPLE SUPPORT DETAILS : CONTINUOUS CARBON FILM SAMPLE VITRIFICATION DETAILS : TANNIC ACID AND GLUCOSE : EMBEDDING AND THEN : LIQUID NITROGEN : FREEZING SAMPLE BUFFER : 60mM MES, pH5.3, : 150mM NaCl, 2.5mM GTP, : 1mM MgSO4, 1mM ZnSO4, : 0.02mg/ml Pepstatin. PH : PH=5.3 SAMPLE DETAILS: NULL DATA ACQUISITION DATE OF EXPERIMENT : 1999-2000 NUMBER OF MICROGRAPHS-IMAGES : NULL TEMPERATURE (KELVIN) : 100.00 MICROSCOPE MODEL : JEOL 4000EX DETECTOR TYPE : GATAN 794 MINIMUM DEFOCUS (NM) : NULL MAXIMUM DEFOCUS (NM) : NULL MINIMUM TILT ANGLE (DEGREES) : 15.00 MAXIMUM TILT ANGLE (DEGREES) : 55.00 NOMINAL CS : NULL IMAGING MODE : DIFFRACTION ELECTRON DOSE (ELECTRONS NM**-2) : 1000.00 ILLUMINATION MODE : LOW-DOSE NOMINAL MAGNIFICATION : NULL CAMERA LENGTH : 150 CM CALIBRATED MAGNIFICATION : NULL SOURCE : LaB6 ACCELERATION VOLTAGE (KV) : 400 IMAGING DETAILS: A WEAK ELECTRON BEAM AND LONG EXPOSURE TIME (40-60S) WERE USED TO MINIMIZE THE VERTICAL BLOOMING STREAK IN THE DIFFRACTION PATTERN RECORDED WITH THE CCD CAMERA. A PATENT APPLICATION IS PENDING WITH RESPECT TO THESE COORDINATES. CONTACT EMORY UNIVERSITY OFFICE OF TECHNOLOGY TRANSFER FOR COMMERCIAL APPLICATIONS OR LICENSING OPPORTUNIES HTTP://WWW.OTT.EMORY.EDU CONTACT JHN FOR QUESTIONS REGARDING COORDINATES AND PROCESSING JHN@WELLYES.COM | ||||||
Remark 400 | COMPOUND THE MODEL OF THE A,B-TUBULIN/EPOTHILONE A COMPLEX WAS DERIVED USING HIGH RESOLUTION ... COMPOUND THE MODEL OF THE A,B-TUBULIN/EPOTHILONE A COMPLEX WAS DERIVED USING HIGH RESOLUTION ELECTRON DIFFRACTIONS FROM TWO DIMENSIONAL CRYSTALS OF TUBULIN INDUCED BY THE PRESENCE OF ZN++ IONS. DEPOSITED ARE COORDINATES FOR EPOTHILONE A BOUND TO AB-TUBULIN DIMER IN THE ZINC-INDUCED SHEETS. THE LIGAND MODEL WAS FIT INTO A DENSITY MAP FOR WHICH THE RESOLUTION IN THE PLANE OF THE SHEET WAS 2.89 ANGSTROMS AND THAT PERPENDICULAR TO THE SHEET WAS ABOUT 4.2 ANGSTROMS AS DESCRIBED IN THE SUPPLEMENTAL TEXT. PROTEIN MODEL HAS NOT BEEN OPTIMIZED AT THE RESOLUTION REPORTED IN REMARK 3 - SEE WWW.ORGANIC.EMORY.EDU/EPO R FREE REPORTED BY THE SOFTWARE ABOVE IS NOT RELEVANT. PHASES WERE DERIVED FROM A PREVIOUS MODEL OF ALPHA/BETA TUBULIN COMPLEXED WITH TAXOL (PDB ID 1JFF). "SHAKING", HIGH TEMPERATURE ANNEALING, AND MODEL AVERAGING WERE COMBINED TO PRODUCE AN OMIT MAP OF THE BOUND EPOTHILONE THAT MINIMIZED SYSTEMATIC BIAS. A NUMBER OF CONFORMATIONAL MODELS WERE FLEXIBLY FITTED INTO THE INITIAL MAP AND TESTED AGAINST THE DIFFRACTIONS BY DIFFERENCE MAP REFINEMENT AS DESCRIBED IN THE PRIMARY REFERENCE. REFINEMENT TO THE FINAL STRUCTURE WAS LIMITED WITHIN AN 8 A RADIUS OF THE LIGAND. ALTHOUGH THE REST OF THE PROTEIN SHOWS LITTLE DEVIATION FROM THAT SEEN IN 1JFF, CERTAIN RESIDUES DO NOT FULLY CONFORM TO RAMACHANDRAN CHARACTERISTICS AND SHOULD BE REGARDED WITH CAUTION. THE FINAL MAPS ASSOCIATED WITH THE PRESENT MODEL WERE DERIVED BY RIGID FITTING OF THE MODELED COMPLEX. AS SUCH, FREE R REPORTED BY THE SOFTWARE ABOVE IS NOT RELEVANT. |
-構造の表示
ムービー |
ムービービューア |
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構造ビューア | 分子: MolmilJmol/JSmol |
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 1tvk.cif.gz | 163.1 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb1tvk.ent.gz | 118.8 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 1tvk.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
文書・要旨 | 1tvk_validation.pdf.gz | 657.3 KB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 1tvk_full_validation.pdf.gz | 776.3 KB | 表示 | |
XML形式データ | 1tvk_validation.xml.gz | 43.7 KB | 表示 | |
CIF形式データ | 1tvk_validation.cif.gz | 59.8 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/tv/1tvk ftp://data.pdbj.org/pub/pdb/validation_reports/tv/1tvk | HTTPS FTP |
-関連構造データ
関連構造データ | 1jffS S: 精密化の開始モデル |
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類似構造データ |
-リンク
-集合体
登録構造単位 |
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1 |
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単位格子 |
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-要素
#1: タンパク質 | 分子量: 48869.117 Da / 分子数: 1 / 由来タイプ: 天然 / 詳細: see REMARK 400 / 由来: (天然) Bos taurus (ウシ) / 器官: Brain / 参照: UniProt: P02550, UniProt: Q2HJ86*PLUS |
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#2: タンパク質 | 分子量: 47940.945 Da / 分子数: 1 / 由来タイプ: 天然 / 詳細: see REMARK 400 / 由来: (天然) Bos taurus (ウシ) / 器官: Brain / 参照: UniProt: P02554, UniProt: Q6B856*PLUS |
#3: 化合物 | ChemComp-GTP / |
#4: 化合物 | ChemComp-GDP / |
#5: 化合物 | ChemComp-EP / |
-実験情報
-実験
実験 | 手法: 電子線結晶学 |
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EM実験 | 試料の集合状態: 2D ARRAY / 3次元再構成法: 電子線結晶学 |
-試料調製
構成要素 | 名称: tubulin 2D crystal / タイプ: COMPLEX |
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試料 | 包埋: YES / シャドウイング: NO / 染色: NO / 凍結: YES |
結晶 | 溶媒含有率: 66.11 % |
-データ収集
顕微鏡 | モデル: JEOL 4000EX 詳細: A weak electron beam and long exposure time (40-60s) were used to minimize the vertical blooming streak in the diffraction pattern recorded with the CCD camera. |
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電子銃 | 加速電圧: 400 kV / 照射モード: SPOT SCAN |
電子レンズ | モード: DIFFRACTION |
試料ホルダ | 温度: 100 K / 傾斜角・最大: 55 ° / 傾斜角・最小: 15 ° |
撮影 | 電子線照射量: 100 e/Å2 / フィルム・検出器のモデル: GENERIC GATAN / 詳細: 2K CCD |
放射 | プロトコル: SINGLE WAVELENGTH / 単色(M)・ラウエ(L): M / 散乱光タイプ: electron |
放射波長 | 相対比: 1 |
-解析
EMソフトウェア |
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3次元再構成 | 解像度: 2.89 Å / 対称性のタイプ: 2D CRYSTAL | |||||||||||||||||||||||||
原子モデル構築 | プロトコル: RIGID BODY FIT / 空間: RECIPROCAL Target criteria: Analysis of comparitive difference densities 詳細: METHOD--annealing, rigid refining REFINEMENT PROTOCOL--rigid body DETAILS--THE MODEL WAS DERIVED USING HIGH RESOLUTION ELECTRON DIFFRACTIONS FROM TWO DIMENSIONAL CRYSTALS OF TUBULIN INDUCED ...詳細: METHOD--annealing, rigid refining REFINEMENT PROTOCOL--rigid body DETAILS--THE MODEL WAS DERIVED USING HIGH RESOLUTION ELECTRON DIFFRACTIONS FROM TWO DIMENSIONAL CRYSTALS OF TUBULIN INDUCED BY THE PRESENCE OF ZN++ IONS. WHAT FOLLOWS ARE THE COORDINATES FOR EPOLTHILONE-A BOUND TO AB-TUBULIN DIMER IN THE ZINC-INDUCED SHEETS. THE LIGAND MODEL WAS FIT INTO A DENSITY MAP FOR WHICH THE RESOLUTION IN THE PLANE OF THE SHEET WAS 2.89 ANGSTROMS AND THAT PERPENDICULAR TO THE SHEET WAS ABOUT 4.2 ANGSTROMS AS DESCRIBED IN THE SUPPLEMENTARY MATERIAL. PHASES WERE DERIVED FROM A PREVIOUS MODEL OF ALPHA/BETA TUBULIN COMPLEXED WITH TAXOL (1JFF). SHAKING, HIGH TEMPERATURE ANANEALING, AND MODEL WERE COMBINED TO PRODUCE AN OMIT MAP OF THE BOUND EPOTHILONE THAT SYSTEMATIC BIAS.A NUMBER OF CONFORMATIONAL MODELS WERE FLEXIBLY FITTED INITIAL MAP AND TESTED AGAINST THE DIFFRACTIONS BY DIFFERENCE MAP AS DESCRIBED IN THE PRIMARY REFERENCE. REFINEMENT TO THE FINAL STAGE WAS LIMITED WITHIN AN 8A RADIUS OF THE LIGAND. ALTHOUGH THE REMAINING PART OF THE PROTEIN SHOWS LITTLE DEVIATION FROM THAT SEEN IN 1JFF, CER NOT FULLY CONFORM WITH RAMACHANDRAN CHARACTERISTICS. AND SHOULD B THE FINAL MAPS ASSOCIATED WITH THE PRESENT MODEL WERE DERIVED BY COMPLEX. AS SUCH, FREE R REPORTED BY THE SOFTWARE BELOW IS NOT RE | |||||||||||||||||||||||||
原子モデル構築 | PDB-ID: 1JFF | |||||||||||||||||||||||||
精密化 | 開始モデル: PDB ID 1JFF 解像度: 2.89→91.29 Å / SU ML: 0.846 / SU R Cruickshank DPI: 0.846 / SU Rfree: 0.861 / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R: 0.675 / ESU R Free: 0.651 / 立体化学のターゲット値: Engh & Huber / 詳細: SEE REMARK 400, COMPOUND
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原子変位パラメータ | Biso mean: 79.8 Å2
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精密化ステップ | サイクル: LAST / 解像度: 2.89→91.29 Å
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LS精密化 シェル | 解像度: 2.89→3.046 Å / Total num. of bins used: 10 /
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