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基本情報
登録情報 | データベース: EMDB / ID: EMD-25046 | |||||||||
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タイトル | Tomogram of mPCDH15/39G7-AuNP complex | |||||||||
![]() | Tomogram of recombinant mPCDH15 extracellular domain in complex with a AuNP/39G7-Fab' conjugate | |||||||||
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![]() | cadherin / antibody / hearing / mechanotransduction / CELL ADHESION | |||||||||
生物種 | ![]() ![]() ![]() ![]() | |||||||||
手法 | 電子線トモグラフィー法 / クライオ電子顕微鏡法 | |||||||||
![]() | Elferich J / Clark S / Ge J / Goehring A / Matsui A / Gouaux E | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Molecular structures and conformations of protocadherin-15 and its complexes on stereocilia elucidated by cryo-electron tomography. 著者: Johannes Elferich / Sarah Clark / Jingpeng Ge / April Goehring / Aya Matsui / Eric Gouaux / ![]() 要旨: Mechanosensory transduction (MT), the conversion of mechanical stimuli into electrical signals, underpins hearing and balance and is carried out within hair cells in the inner ear. Hair cells harbor ...Mechanosensory transduction (MT), the conversion of mechanical stimuli into electrical signals, underpins hearing and balance and is carried out within hair cells in the inner ear. Hair cells harbor actin-filled stereocilia, arranged in rows of descending heights, where the tips of stereocilia are connected to their taller neighbors by a filament composed of protocadherin 15 (PCDH15) and cadherin 23 (CDH23), deemed the 'tip link.' Tension exerted on the tip link opens an ion channel at the tip of the shorter stereocilia, thus converting mechanical force into an electrical signal. While biochemical and structural studies have provided insights into the molecular composition and structure of isolated portions of the tip link, the architecture, location, and conformational states of intact tip links, on stereocilia, remains unknown. Here, we report in situ cryo-electron microscopy imaging of the tip link in mouse stereocilia. We observe individual PCDH15 molecules at the tip and shaft of stereocilia and determine their stoichiometry, conformational heterogeneity, and their complexes with other filamentous proteins, perhaps including CDH23. The PCDH15 complexes occur in clusters, frequently with more than one copy of PCDH15 at the tip of stereocilia, suggesting that tip links might consist of more than one copy of PCDH15 complexes and, by extension, might include multiple MT complexes. #1: ![]() タイトル: Molecular structure and conformation of stereocilia tip-links elucidated by cryo-electron tomography 著者: Elferich J / Clark S / Ge J / Goehring A / Matsui A / Gouaux E | |||||||||
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構造の表示
ムービー |
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添付画像 |
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-EMDBアーカイブ
マップデータ | ![]() | 172.6 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 11.9 KB 11.9 KB | 表示 表示 | ![]() |
画像 | ![]() | 151 KB | ||
Filedesc metadata | ![]() | 3.7 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 405.5 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 405 KB | 表示 | |
XML形式データ | ![]() | 4.8 KB | 表示 | |
CIF形式データ | ![]() | 5.8 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | C: 同じ文献を引用 ( |
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電子顕微鏡画像生データ | ![]() Data size: 58.5 Data #1: Tiltseries of mPCDH15/39G7-AuNP complex [tilt series] Data #2: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #3: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #4: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #5: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #6: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #7: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #8: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #9: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #10: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #11: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #12: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #13: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #14: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #15: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #16: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series]) ![]() Data size: 1.9 TB Data #1: Unaligned tilt series of mouse stereocilia [tilt series]) |
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リンク
EMDBのページ | ![]() ![]() |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Tomogram of recombinant mPCDH15 extracellular domain in complex with a AuNP/39G7-Fab' conjugate | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 5.756 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 : mPCDH15/39G7-AuNP complex
全体 | 名称: mPCDH15/39G7-AuNP complex |
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要素 |
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-超分子 #1: mPCDH15/39G7-AuNP complex
超分子 | 名称: mPCDH15/39G7-AuNP complex / タイプ: complex / ID: 1 / 親要素: 0 |
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-超分子 #2: 39G7-AuNP conjugate
超分子 | 名称: 39G7-AuNP conjugate / タイプ: complex / ID: 2 / 親要素: 1 |
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由来(天然) | 生物種: ![]() ![]() |
-超分子 #3: mPCDH15 extracellular domain
超分子 | 名称: mPCDH15 extracellular domain / タイプ: complex / ID: 3 / 親要素: 1 |
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由来(天然) | 生物種: ![]() ![]() |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 電子線トモグラフィー法 |
試料の集合状態 | particle |
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試料調製
緩衝液 | pH: 7.4 |
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凍結 | 凍結剤: ETHANE |
切片作成 | その他: NO SECTIONING |
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電子顕微鏡法
顕微鏡 | FEI TALOS ARCTICA |
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撮影 | フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 実像数: 41 / 平均電子線量: 3.0 e/Å2 |
電子線 | 加速電圧: 200 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm |
実験機器 | ![]() モデル: Talos Arctica / 画像提供: FEI Company |
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画像解析
最終 再構成 | アルゴリズム: BACK PROJECTION / ソフトウェア - 名称: ![]() |
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