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基本情報
登録情報 | データベース: EMDB / ID: EMD-4154 | |||||||||||||||||||||
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タイトル | Mechanism of microtubule minus-end recognition and protection by CAMSAP proteins | |||||||||||||||||||||
![]() | CAMSAP3 CKK decorated 13pf microtubule asymmetric unit | |||||||||||||||||||||
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![]() | CAMSAP CKK Microtubule Tubulin / Structural protein | |||||||||||||||||||||
機能・相同性 | ![]() regulation of organelle organization / zonula adherens maintenance / microtubule minus-end / protein transport along microtubule / microtubule anchoring / 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 ...regulation of organelle organization / zonula adherens maintenance / microtubule minus-end / protein transport along microtubule / microtubule anchoring / 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 / microtubule minus-end binding / RHO GTPases Activate Formins / regulation of Golgi organization / establishment or maintenance of microtubule cytoskeleton polarity / MHC class II antigen presentation / cilium movement / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / Aggrephagy / epithelial cell-cell adhesion / The role of GTSE1 in G2/M progression after G2 checkpoint / zonula adherens / 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 / Recruitment of NuMA to mitotic centrosomes / Regulation of PLK1 Activity at G2/M Transition / Hedgehog 'off' state / negative regulation of microtubule depolymerization / establishment of epithelial cell apical/basal polarity / motile cilium / positive regulation of axon guidance / embryo development ending in birth or egg hatching / regulation of focal adhesion assembly / spectrin binding / regulation of microtubule polymerization / axoneme / microtubule-based process / regulation of microtubule cytoskeleton organization / cytoplasmic microtubule organization / regulation of cell migration / ciliary basal body / 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 / neuron projection development / actin filament binding / mitotic cell cycle / nervous system development / in utero embryonic development / microtubule / calmodulin binding / hydrolase activity / protein heterodimerization activity / GTPase activity / centrosome / GTP binding / metal ion binding / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||||||||||||||
生物種 | ![]() ![]() ![]() ![]() | |||||||||||||||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 5.3 Å | |||||||||||||||||||||
![]() | Akhmanova A / Moores CA | |||||||||||||||||||||
資金援助 | ![]() ![]() ![]() ![]()
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![]() | ![]() タイトル: A structural model for microtubule minus-end recognition and protection by CAMSAP proteins. 著者: Joseph Atherton / Kai Jiang / Marcel M Stangier / Yanzhang Luo / Shasha Hua / Klaartje Houben / Jolien J E van Hooff / Agnel-Praveen Joseph / Guido Scarabelli / Barry J Grant / Anthony J ...著者: Joseph Atherton / Kai Jiang / Marcel M Stangier / Yanzhang Luo / Shasha Hua / Klaartje Houben / Jolien J E van Hooff / Agnel-Praveen Joseph / Guido Scarabelli / Barry J Grant / Anthony J Roberts / Maya Topf / Michel O Steinmetz / Marc Baldus / Carolyn A Moores / Anna Akhmanova / ![]() ![]() ![]() ![]() 要旨: CAMSAP and Patronin family members regulate microtubule minus-end stability and localization and thus organize noncentrosomal microtubule networks, which are essential for cell division, polarization ...CAMSAP and Patronin family members regulate microtubule minus-end stability and localization and thus organize noncentrosomal microtubule networks, which are essential for cell division, polarization and differentiation. Here, we found that the CAMSAP C-terminal CKK domain is widely present among eukaryotes and autonomously recognizes microtubule minus ends. Through a combination of structural approaches, we uncovered how mammalian CKK binds between two tubulin dimers at the interprotofilament interface on the outer microtubule surface. In vitro reconstitution assays combined with high-resolution fluorescence microscopy and cryo-electron tomography suggested that CKK preferentially associates with the transition zone between curved protofilaments and the regular microtubule lattice. We propose that minus-end-specific features of the interprotofilament interface at this site serve as the basis for CKK's minus-end preference. The steric clash between microtubule-bound CKK and kinesin motors explains how CKK protects microtubule minus ends against kinesin-13-induced depolymerization and thus controls the stability of free microtubule minus ends. | |||||||||||||||||||||
履歴 |
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構造の表示
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構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
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マップデータ | ![]() | 715.7 KB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 17.4 KB 17.4 KB | 表示 表示 | ![]() |
画像 | ![]() | 269.5 KB | ||
Filedesc metadata | ![]() | 6.8 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
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リンク
EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | CAMSAP3 CKK decorated 13pf microtubule asymmetric unit | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.534 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
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試料の構成要素
+全体 : Complex of two tubulin dimers with bound CAMSAP3-CKK domain
+超分子 #1: Complex of two tubulin dimers with bound CAMSAP3-CKK domain
+超分子 #2: tubulin
+超分子 #3: CAMSAP3-CKK domain
+分子 #1: Tubulin alpha chain
+分子 #2: Tubulin beta-2B chain
+分子 #3: Calmodulin-regulated spectrin-associated protein 3
+分子 #4: GUANOSINE-5'-TRIPHOSPHATE
+分子 #5: MAGNESIUM ION
+分子 #6: GUANOSINE-5'-DIPHOSPHATE
+分子 #7: TAXOL
+分子 #8: water
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | filament |
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試料調製
緩衝液 | pH: 6.8 / 詳細: BRB80 |
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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 |
凍結 | 凍結剤: ETHANE / 装置: FEI VITROBOT MARK III |
詳細 | 13pf Microtubules |
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電子顕微鏡法
顕微鏡 | FEI POLARA 300 |
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撮影 | フィルム・検出器のモデル: DIRECT ELECTRON DE-20 (5k x 3k) 検出モード: INTEGRATING / 平均電子線量: 25.0 e/Å2 / 詳細: 25e-/A2 used in final reconstuctions |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD |
実験機器 | ![]() モデル: Tecnai Polara / 画像提供: FEI Company |
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画像解析
初期モデル | モデルのタイプ: OTHER / 詳細: Kinesin decorated microtubule |
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最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 5.3 Å / 解像度の算出法: FSC 0.143 CUT-OFF 詳細: Gold-standard noise substitution test used to asses for over fitting (Chen et al., 2013) 使用した粒子像数: 6530 |
初期 角度割当 | タイプ: PROJECTION MATCHING / ソフトウェア - 名称: SPIDER |
最終 角度割当 | タイプ: OTHER / ソフトウェア - 名称: FREALIGN / 詳細: Projection matching in Fourier space |
-原子モデル構築 1
精密化 | プロトコル: OTHER |
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得られたモデル | ![]() PDB-5m50: |