+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-19042 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
タイトル | MAP7 MTBD (microtubule binding domain) decorated microtubule protofilament | |||||||||
マップデータ | Unsharpened map of MAP7 MTBD bound microtubule protofilament | |||||||||
試料 |
| |||||||||
キーワード | Microtubule associated protein 7(MAP 7) / microtubule binding domain / cryo-EM / STRUCTURAL PROTEIN | |||||||||
機能・相同性 | 機能・相同性情報 Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Hedgehog 'off' state / Cilium Assembly / Intraflagellar transport / COPI-dependent Golgi-to-ER retrograde traffic / Carboxyterminal post-translational modifications of tubulin / RHOH GTPase cycle / Sealing of the nuclear envelope (NE) by ESCRT-III / Kinesins / PKR-mediated signaling ...Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Hedgehog 'off' state / Cilium Assembly / Intraflagellar transport / COPI-dependent Golgi-to-ER retrograde traffic / Carboxyterminal post-translational modifications of tubulin / RHOH GTPase cycle / Sealing of the nuclear envelope (NE) by ESCRT-III / Kinesins / PKR-mediated signaling / The role of GTSE1 in G2/M progression after G2 checkpoint / Aggrephagy / Resolution of Sister Chromatid Cohesion / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / Separation of Sister Chromatids / RHO GTPases activate IQGAPs / RHO GTPases Activate Formins / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / MHC class II antigen presentation / Recruitment of NuMA to mitotic centrosomes / COPI-mediated anterograde transport / structural constituent of cytoskeleton / microtubule cytoskeleton organization / microtubule cytoskeleton / mitotic cell cycle / 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 / microtubule / GTPase activity / GTP binding / metal ion binding / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) / Sus scrofa (ブタ) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.7 Å | |||||||||
データ登録者 | Bangera M / Moores CA | |||||||||
資金援助 | 英国, 1件
| |||||||||
引用 | ジャーナル: Nat Commun / 年: 2024 タイトル: A structural and dynamic visualization of the interaction between MAP7 and microtubules. 著者: Agnes Adler / Mamata Bangera / J Wouter Beugelink / Salima Bahri / Hugo van Ingen / Carolyn A Moores / Marc Baldus / 要旨: Microtubules (MTs) are key components of the eukaryotic cytoskeleton and are essential for intracellular organization, organelle trafficking and mitosis. MT tasks depend on binding and interactions ...Microtubules (MTs) are key components of the eukaryotic cytoskeleton and are essential for intracellular organization, organelle trafficking and mitosis. MT tasks depend on binding and interactions with MT-associated proteins (MAPs). MT-associated protein 7 (MAP7) has the unusual ability of both MT binding and activating kinesin-1-mediated cargo transport along MTs. Additionally, the protein is reported to stabilize MTs with its 112 amino-acid long MT-binding domain (MTBD). Here we investigate the structural basis of the interaction of MAP7 MTBD with the MT lattice. Using a combination of solid and solution-state nuclear magnetic resonance (NMR) spectroscopy with electron microscopy, fluorescence anisotropy and isothermal titration calorimetry, we shed light on the binding mode of MAP7 to MTs at an atomic level. Our results show that a combination of interactions between MAP7 and MT lattice extending beyond a single tubulin dimer and including tubulin C-terminal tails contribute to formation of the MAP7-MT complex. | |||||||||
履歴 |
|
-構造の表示
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_19042.map.gz | 170.7 MB | EMDBマップデータ形式 | |
---|---|---|---|---|
ヘッダ (付随情報) | emd-19042-v30.xml emd-19042.xml | 24.9 KB 24.9 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_19042_fsc.xml | 14.9 KB | 表示 | FSCデータファイル |
画像 | emd_19042.png | 47.5 KB | ||
Filedesc metadata | emd-19042.cif.gz | 7.4 KB | ||
その他 | emd_19042_additional_1.map.gz emd_19042_additional_2.map.gz emd_19042_half_map_1.map.gz emd_19042_half_map_2.map.gz | 270.8 MB 192.8 MB 317.8 MB 317.8 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-19042 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-19042 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_19042_validation.pdf.gz | 1.2 MB | 表示 | EMDB検証レポート |
---|---|---|---|---|
文書・詳細版 | emd_19042_full_validation.pdf.gz | 1.2 MB | 表示 | |
XML形式データ | emd_19042_validation.xml.gz | 24.2 KB | 表示 | |
CIF形式データ | emd_19042_validation.cif.gz | 31.6 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-19042 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-19042 | HTTPS FTP |
-関連構造データ
関連構造データ | 8rc1MC M: このマップから作成された原子モデル C: 同じ文献を引用 (文献) |
---|---|
類似構造データ | 類似検索 - 機能・相同性F&H 検索 |
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
---|---|
「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_19042.map.gz / 形式: CCP4 / 大きさ: 343 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Unsharpened map of MAP7 MTBD bound microtubule protofilament | ||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.067 Å | ||||||||||||||||||||||||||||||||||||
密度 |
| ||||||||||||||||||||||||||||||||||||
対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
|
-添付データ
-追加マップ: Unsharpened C1 reconstruction of MAP7 MTBD bound microtubule
ファイル | emd_19042_additional_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Unsharpened C1 reconstruction of MAP7 MTBD bound microtubule | ||||||||||||
投影像・断面図 |
| ||||||||||||
密度ヒストグラム |
-追加マップ: Local resolution filtered symmetrized reconstruction of MAP7 MTBD...
ファイル | emd_19042_additional_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Local resolution filtered symmetrized reconstruction of MAP7 MTBD bound microtubule with an applied B factor of -50 | ||||||||||||
投影像・断面図 |
| ||||||||||||
密度ヒストグラム |
-ハーフマップ: Unsharpened half map 1
ファイル | emd_19042_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Unsharpened half map 1 | ||||||||||||
投影像・断面図 |
| ||||||||||||
密度ヒストグラム |
-ハーフマップ: Unsharpened half map 2
ファイル | emd_19042_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Unsharpened half map 2 | ||||||||||||
投影像・断面図 |
| ||||||||||||
密度ヒストグラム |
-試料の構成要素
-全体 : Complex of MAP7 MTBD (microtubule binding domain) with 14pf micro...
全体 | 名称: Complex of MAP7 MTBD (microtubule binding domain) with 14pf microtubule |
---|---|
要素 |
|
-超分子 #1: Complex of MAP7 MTBD (microtubule binding domain) with 14pf micro...
超分子 | 名称: Complex of MAP7 MTBD (microtubule binding domain) with 14pf microtubule タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#3 |
---|---|
由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 13 kDa/nm |
-分子 #1: Tubulin alpha-1B chain
分子 | 名称: Tubulin alpha-1B chain / タイプ: protein_or_peptide / ID: 1 / コピー数: 2 / 光学異性体: LEVO |
---|---|
由来(天然) | 生物種: Sus scrofa (ブタ) |
分子量 | 理論値: 50.204445 KDa |
配列 | 文字列: MRECISIHVG QAGVQIGNAC WELYCLEHGI QPDGQMPSDK TIGGGDDSFN TFFSETGAGK HVPRAVFVDL EPTVIDEVRT GTYRQLFHP EQLITGKEDA ANNYARGHYT IGKEIIDLVL DRIRKLADQC TGLQGFLVFH SFGGGTGSGF TSLLMERLSV D YGKKSKLE ...文字列: MRECISIHVG QAGVQIGNAC WELYCLEHGI QPDGQMPSDK TIGGGDDSFN TFFSETGAGK HVPRAVFVDL EPTVIDEVRT GTYRQLFHP EQLITGKEDA ANNYARGHYT IGKEIIDLVL DRIRKLADQC TGLQGFLVFH SFGGGTGSGF TSLLMERLSV D YGKKSKLE FSIYPAPQVS TAVVEPYNSI LTTHTTLEHS DCAFMVDNEA IYDICRRNLD IERPTYTNLN RLISQIVSSI TA SLRFDGA LNVDLTEFQT NLVPYPRIHF PLATYAPVIS AEKAYHEQLS VAEITNACFE PANQMVKCDP RHGKYMACCL LYR GDVVPK DVNAAIATIK TKRSIQFVDW CPTGFKVGIN YQPPTVVPGG DLAKVQRAVC MLSNTTAIAE AWARLDHKFD LMYA KRAFV HWYVGEGMEE GEFSEAREDM AALEKDYEEV GVDSVEGEGE EEGEEY UniProtKB: Tubulin alpha-1B chain |
-分子 #2: Tubulin beta chain
分子 | 名称: Tubulin beta chain / タイプ: protein_or_peptide / ID: 2 / コピー数: 2 / 光学異性体: LEVO |
---|---|
由来(天然) | 生物種: Sus scrofa (ブタ) |
分子量 | 理論値: 48.184207 KDa |
配列 | 文字列: MREIVHIQAG QCGNQIGAKF WEVISDEHGI DPTGSYHGDS DLQLERINVY YNEAAGNKYV PRAILVDLEP GTMDSVRSGP FGQIFRPDN FVFGQSGAGN NWAKGHYTEG AELVDSVLDV VRKESESCDC LQGFQLTHSL GGGTGSGMGT LLISKIREEY P DRIMNTFS ...文字列: MREIVHIQAG QCGNQIGAKF WEVISDEHGI DPTGSYHGDS DLQLERINVY YNEAAGNKYV PRAILVDLEP GTMDSVRSGP FGQIFRPDN FVFGQSGAGN NWAKGHYTEG AELVDSVLDV VRKESESCDC LQGFQLTHSL GGGTGSGMGT LLISKIREEY P DRIMNTFS VVPSPKVSDT VVEPYNATLS VHQLVENTDE TYCIDNEALY DICFRTLKLT TPTYGDLNHL VSATMSGVTT CL RFPGQLN ADLRKLAVNM VPFPRLHFFM PGFAPLTSRG SQQYRALTVP ELTQQMFDAK NMMAACDPRH GRYLTVAAVF RGR MSMKEV DEQMLNVQNK NSSYFVEWIP NNVKTAVCDI PPRGLKMSAT FIGNSTAIQE LFKRISEQFT AMFRRKAFLH WYTG EGMDE MEFTEAESNM NDLVSEYQQY QDATA UniProtKB: Tubulin beta chain |
-分子 #3: Microtubule associated protein 7 (MAP7)
分子 | 名称: Microtubule associated protein 7 (MAP7) / タイプ: protein_or_peptide / ID: 3 / コピー数: 1 / 光学異性体: LEVO |
---|---|
由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 7.59235 KDa |
組換発現 | 生物種: Escherichia coli (大腸菌) |
配列 | 文字列: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) ...文字列: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) |
-分子 #4: GUANOSINE-5'-TRIPHOSPHATE
分子 | 名称: GUANOSINE-5'-TRIPHOSPHATE / タイプ: ligand / ID: 4 / コピー数: 2 / 式: GTP |
---|---|
分子量 | 理論値: 523.18 Da |
Chemical component information | ChemComp-GTP: |
-分子 #5: MAGNESIUM ION
分子 | 名称: MAGNESIUM ION / タイプ: ligand / ID: 5 / コピー数: 2 / 式: MG |
---|---|
分子量 | 理論値: 24.305 Da |
-分子 #6: GUANOSINE-5'-DIPHOSPHATE
分子 | 名称: GUANOSINE-5'-DIPHOSPHATE / タイプ: ligand / ID: 6 / コピー数: 2 / 式: GDP |
---|---|
分子量 | 理論値: 443.201 Da |
Chemical component information | ChemComp-GDP: |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
---|---|
解析 | 単粒子再構成法 |
試料の集合状態 | filament |
-試料調製
濃度 | 0.5 mg/mL |
---|---|
緩衝液 | pH: 6.5 / 構成要素 - 名称: BRB80 / 詳細: 80mM PIPES, 2mM MgCl2, 1mM EGTA |
グリッド | モデル: C-flat-2/2 / 材質: COPPER / メッシュ: 400 / 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: PLASMA CLEANING / 前処理 - 時間: 40 sec. / 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 IV 詳細: Microtubules stabilised by MAP7 MTBD were applied to a glow discharged grid, incubated for 30 seconds at room temperature following which excess sample was wicked off and MAP7 MTBD was ...詳細: Microtubules stabilised by MAP7 MTBD were applied to a glow discharged grid, incubated for 30 seconds at room temperature following which excess sample was wicked off and MAP7 MTBD was immediately added to the grid. After incubation for 60 seconds in a Vitrobot, excess liquid was blotted off and the grid was plunge frozen in liquid ethane.. |
詳細 | Equimolar concentrations of tubulin and MAP7 MTBD were mixed with GTP and incubated in a water bath maintained at 37 degrees for 1 hour |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
---|---|
撮影 | フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) 平均電子線量: 49.33 e/Å2 詳細: Movies were collected in counting mode fractionated over 50 frames |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2.4 µm / 最小 デフォーカス(公称値): 0.6 µm / 倍率(公称値): 81000 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
+画像解析
-原子モデル構築 1
初期モデル |
| ||||||
---|---|---|---|---|---|---|---|
得られたモデル | PDB-8rc1: |