+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-3778 | |||||||||
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タイトル | Molecular basis of human kinesin-8 function and inhibition | |||||||||
マップデータ | Asymmetric unit extracted from the full MT-bound Kif18A motor domain no nucleotide reconstruction | |||||||||
試料 |
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機能・相同性 | 機能・相同性情報 tubulin-dependent ATPase activity / mitotic spindle astral microtubule / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Intraflagellar transport / Carboxyterminal post-translational modifications of tubulin / Sealing of the nuclear envelope (NE) by ESCRT-III / Kinesins / Resolution of Sister Chromatid Cohesion / Mitotic Prometaphase ...tubulin-dependent ATPase activity / mitotic spindle astral microtubule / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Intraflagellar transport / Carboxyterminal post-translational modifications of tubulin / Sealing of the nuclear envelope (NE) by ESCRT-III / Kinesins / Resolution of Sister Chromatid Cohesion / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / COPI-dependent Golgi-to-ER retrograde traffic / RHO GTPases activate IQGAPs / COPI-independent Golgi-to-ER retrograde traffic / COPI-mediated anterograde transport / RHO GTPases Activate Formins / MHC class II antigen presentation / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / mitotic spindle midzone / Aggrephagy / kinetochore microtubule / The role of GTSE1 in G2/M progression after G2 checkpoint / Separation of Sister Chromatids / Loss of Nlp from mitotic centrosomes / Recruitment of mitotic centrosome proteins and complexes / Loss of proteins required for interphase microtubule organization from the centrosome / Anchoring of the basal body to the plasma membrane / AURKA Activation by TPX2 / male meiotic nuclear division / Recruitment of NuMA to mitotic centrosomes / Regulation of PLK1 Activity at G2/M Transition / Hedgehog 'off' state / microtubule plus-end binding / Kinesins / plus-end-directed microtubule motor activity / COPI-dependent Golgi-to-ER retrograde traffic / microtubule depolymerization / kinesin complex / mitotic metaphase chromosome alignment / microtubule-based movement / seminiferous tubule development / mitotic sister chromatid segregation / cytoplasmic microtubule / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / regulation of microtubule cytoskeleton organization / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / ruffle / Resolution of Sister Chromatid Cohesion / MHC class II antigen presentation / cellular response to interleukin-4 / cellular response to estradiol stimulus / カベオラ / RHO GTPases Activate Formins / 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 / 動原体 / Separation of Sister Chromatids / microtubule cytoskeleton / protein transport / double-stranded RNA binding / mitotic cell cycle / actin binding / microtubule binding / 微小管 / hydrolase activity / GTPase activity / 中心体 / ubiquitin protein ligase binding / GTP binding / ATP hydrolysis activity / ATP binding / metal ion binding / 細胞核 / 細胞質基質 / 細胞質 類似検索 - 分子機能 | |||||||||
生物種 | Bos taurus (ウシ) / Sus scrofa (ブタ) / Homo sapiens (ヒト) / Bovine (ウシ) / Pig (ブタ) | |||||||||
手法 | らせん対称体再構成法 / クライオ電子顕微鏡法 / 解像度: 5.5 Å | |||||||||
データ登録者 | Locke J / Joseph AP / Topf M / Moores CA | |||||||||
資金援助 | 英国, 2件
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引用 | ジャーナル: Proc Natl Acad Sci U S A / 年: 2017 タイトル: Structural basis of human kinesin-8 function and inhibition. 著者: Julia Locke / Agnel Praveen Joseph / Alejandro Peña / Martin M Möckel / Thomas U Mayer / Maya Topf / Carolyn A Moores / 要旨: Kinesin motors play diverse roles in mitosis and are targets for antimitotic drugs. The clinical significance of these motors emphasizes the importance of understanding the molecular basis of their ...Kinesin motors play diverse roles in mitosis and are targets for antimitotic drugs. The clinical significance of these motors emphasizes the importance of understanding the molecular basis of their function. Equally important, investigations into the modes of inhibition of these motors provide crucial information about their molecular mechanisms. Kif18A regulates spindle microtubules through its dual functionality, with microtubule-based stepping and regulation of microtubule dynamics. We investigated the mechanism of Kif18A and its inhibition by the small molecule BTB-1. The Kif18A motor domain drives ATP-dependent plus-end microtubule gliding, and undergoes conformational changes consistent with canonical mechanisms of plus-end-directed motility. The Kif18A motor domain also depolymerizes microtubule plus and minus ends. BTB-1 inhibits both of these microtubule-based Kif18A activities. A reconstruction of BTB-1-bound, microtubule-bound Kif18A, in combination with computational modeling, identified an allosteric BTB-1-binding site near loop5, where it blocks the ATP-dependent conformational changes that we characterized. Strikingly, BTB-1 binding is close to that of well-characterized Kif11 inhibitors that block tight microtubule binding, whereas BTB-1 traps Kif18A on the microtubule. Our work highlights a general mechanism of kinesin inhibition in which small-molecule binding near loop5 prevents a range of conformational changes, blocking motor function. | |||||||||
履歴 |
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-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 | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_3778.map.gz | 452.4 KB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-3778-v30.xml emd-3778.xml | 18.9 KB 18.9 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_3778_fsc.xml | 17.8 KB | 表示 | FSCデータファイル |
画像 | emd_3778.png | 65.8 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-3778 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-3778 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_3778.map.gz / 形式: CCP4 / 大きさ: 1.3 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Asymmetric unit extracted from the full MT-bound Kif18A motor domain no nucleotide reconstruction | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.39 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
-試料の構成要素
+全体 : 13-Protofilament microtuble-bound human Kinesin-8 motor domain ne...
+超分子 #1: 13-Protofilament microtuble-bound human Kinesin-8 motor domain ne...
+超分子 #2: Tubulin, alpha
+超分子 #3: Tublin, beta
+超分子 #4: Kinesin-8 motor domain neck-linker
+分子 #1: Kinesin-like protein KIF18A
+分子 #2: Tubulin alpha chain
+分子 #3: Tubulin beta chain
+分子 #4: ZINC ION
+分子 #5: MAGNESIUM ION
+分子 #6: GUANOSINE-5'-TRIPHOSPHATE
+分子 #7: GUANOSINE-5'-DIPHOSPHATE
+分子 #8: TAXOL
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | らせん対称体再構成法 |
試料の集合状態 | filament |
-試料調製
緩衝液 | pH: 7.4 |
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グリッド | モデル: C-flat-2/2 / 材質: COPPER / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 雰囲気: AIR |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 298 K / 装置: FEI VITROBOT MARK III |
-電子顕微鏡法
顕微鏡 | FEI POLARA 300 |
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電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 最大 デフォーカス(補正後): 3.5 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELDBright-field microscopy / Cs: 2.3 mm / 最小 デフォーカス(公称値): 0.5 µm |
特殊光学系 | エネルギーフィルター - 名称: GIF Quantum |
試料ステージ | 試料ホルダーモデル: OTHER / ホルダー冷却材: NITROGEN |
詳細 | FEI Polara |
撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: COUNTING / デジタル化 - サイズ - 横: 3840 pixel / デジタル化 - サイズ - 縦: 3712 pixel / 平均露光時間: 2.0 sec. / 平均電子線量: 5.0 e/Å2 |
実験機器 | モデル: Tecnai Polara / 画像提供: FEI Company |
-画像解析
-原子モデル構築 1
精密化 | 空間: REAL / プロトコル: FLEXIBLE FIT |
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得られたモデル | PDB-5oam: |