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基本情報
登録情報 | データベース: EMDB / ID: EMD-3103 | |||||||||
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タイトル | entire human nuclear pore complex | |||||||||
![]() | Reconstruction of the human nuclear pore complex | |||||||||
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機能・相同性 | ![]() GATOR2 complex / nephron development / Seh1-associated complex / nuclear pore inner ring / protein localization to nuclear inner membrane / protein exit from endoplasmic reticulum / COPII-coated vesicle budding / nuclear envelope organization / telomere tethering at nuclear periphery / COPII-coated vesicle cargo loading ...GATOR2 complex / nephron development / Seh1-associated complex / nuclear pore inner ring / protein localization to nuclear inner membrane / protein exit from endoplasmic reticulum / COPII-coated vesicle budding / nuclear envelope organization / telomere tethering at nuclear periphery / COPII-coated vesicle cargo loading / transcription-dependent tethering of RNA polymerase II gene DNA at nuclear periphery / nuclear pore organization / nuclear pore outer ring / ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() | |||||||||
手法 | サブトモグラム平均法 / ![]() | |||||||||
![]() | von Appen A / Kosinski J / Sparks L / Ori A / DiGuilio A / Vollmer B / Mackmull M / Banterle N / Parca L / Kastritis P ...von Appen A / Kosinski J / Sparks L / Ori A / DiGuilio A / Vollmer B / Mackmull M / Banterle N / Parca L / Kastritis P / Buczak K / Mosalaganti S / Hagen W / Andres-Pons A / Lemke EA / Bork P / Antonin W / Glavy JS / Bui KH / Beck M | |||||||||
![]() | ![]() タイトル: In situ structural analysis of the human nuclear pore complex. 著者: Alexander von Appen / Jan Kosinski / Lenore Sparks / Alessandro Ori / Amanda L DiGuilio / Benjamin Vollmer / Marie-Therese Mackmull / Niccolo Banterle / Luca Parca / Panagiotis Kastritis / ...著者: Alexander von Appen / Jan Kosinski / Lenore Sparks / Alessandro Ori / Amanda L DiGuilio / Benjamin Vollmer / Marie-Therese Mackmull / Niccolo Banterle / Luca Parca / Panagiotis Kastritis / Katarzyna Buczak / Shyamal Mosalaganti / Wim Hagen / Amparo Andres-Pons / Edward A Lemke / Peer Bork / Wolfram Antonin / Joseph S Glavy / Khanh Huy Bui / Martin Beck / ![]() ![]() ![]() 要旨: Nuclear pore complexes are fundamental components of all eukaryotic cells that mediate nucleocytoplasmic exchange. Determining their 110-megadalton structure imposes a formidable challenge and ...Nuclear pore complexes are fundamental components of all eukaryotic cells that mediate nucleocytoplasmic exchange. Determining their 110-megadalton structure imposes a formidable challenge and requires in situ structural biology approaches. Of approximately 30 nucleoporins (Nups), 15 are structured and form the Y and inner-ring complexes. These two major scaffolding modules assemble in multiple copies into an eight-fold rotationally symmetric structure that fuses the inner and outer nuclear membranes to form a central channel of ~60 nm in diameter. The scaffold is decorated with transport-channel Nups that often contain phenylalanine-repeat sequences and mediate the interaction with cargo complexes. Although the architectural arrangement of parts of the Y complex has been elucidated, it is unclear how exactly it oligomerizes in situ. Here we combine cryo-electron tomography with mass spectrometry, biochemical analysis, perturbation experiments and structural modelling to generate, to our knowledge, the most comprehensive architectural model of the human nuclear pore complex to date. Our data suggest previously unknown protein interfaces across Y complexes and to inner-ring complex members. We show that the transport-channel Nup358 (also known as Ranbp2) has a previously unanticipated role in Y-complex oligomerization. Our findings blur the established boundaries between scaffold and transport-channel Nups. We conclude that, similar to coated vesicles, several copies of the same structural building block--although compositionally identical--engage in different local sets of interactions and conformations. | |||||||||
履歴 |
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構造の表示
ムービー |
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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 | ![]() | 13.9 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 10.1 KB 10.1 KB | 表示 表示 | ![]() |
画像 | ![]() | 200.7 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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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 | Reconstruction of the human nuclear pore complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 6.84 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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試料の構成要素
-全体 : Human Nuclear Pore Complex
全体 | 名称: Human Nuclear Pore Complex![]() |
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要素 |
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-超分子 #1000: Human Nuclear Pore Complex
超分子 | 名称: Human Nuclear Pore Complex / タイプ: sample / ID: 1000 詳細: The sample is purified human nuclear envelope containing nuclear pore complex. Number unique components: 1 |
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分子量 | 手法: Absolute Quantitative Mass Spectrometry |
-分子 #1: Nuclear Pore Complex
分子 | 名称: Nuclear Pore Complex / タイプ: protein_or_peptide / ID: 1 / コピー数: 1 / 組換発現: No |
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由来(天然) | 生物種: ![]() ![]() |
-実験情報
-構造解析
手法 | ![]() |
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![]() | サブトモグラム平均法 |
試料の集合状態 | particle |
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試料調製
緩衝液 | pH: 7.5 / 詳細: 20mM Tris, 0.2-0.4% Trehalose |
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グリッド | 詳細: 200 mesh Quantifoil Copper Holey Carbon Grid R2/1 |
凍結 | 凍結剤: ETHANE-PROPANE MIXTURE / 装置: HOMEMADE PLUNGER |
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電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 倍率(補正後): 42000 / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD![]() |
特殊光学系 | エネルギーフィルター - 名称: GIF Quantum エネルギーフィルター - エネルギー下限: 0.0 eV エネルギーフィルター - エネルギー上限: 20.0 eV |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER Tilt series - Axis1 - Min angle: -45 ° / Tilt series - Axis1 - Max angle: 60 ° |
日付 | 2014年10月17日 |
撮影 | カテゴリ: CCD / フィルム・検出器のモデル: GATAN K2 (4k x 4k) / 平均電子線量: 110 e/Å2 / ビット/ピクセル: 16 |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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画像解析
CTF補正 | 詳細: Phase flipping of tilt series |
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最終 再構成 | 想定した対称性 - 点群: C8 (8回回転対称![]() |
詳細 | The subtomograms were picked manually and further processed iterative symmetry independent averaging. |