+
データを開く
-
基本情報
登録情報 | ![]() | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
タイトル | 48-nm repeating structure of doublets from mouse sperm flagella | ||||||||||||
![]() | Final map of 48 nm-repeating unit of mouse sperm doublets | ||||||||||||
![]() |
| ||||||||||||
![]() | Mammalian sperm / axoneme / microtubule-based structure / microtubule inner protein / non-motor proteins / cellular motility / fertility / structural protein | ||||||||||||
機能・相同性 | ![]() ERKs are inactivated / 9+0 motile cilium / sperm flagellum assembly / outer acrosomal membrane / regulation of brood size / establishment of left/right asymmetry / protein localization to motile cilium / manchette assembly / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly ...ERKs are inactivated / 9+0 motile cilium / sperm flagellum assembly / outer acrosomal membrane / regulation of brood size / establishment of left/right asymmetry / protein localization to motile cilium / manchette assembly / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Sealing of the nuclear envelope (NE) by ESCRT-III / left/right pattern formation / axonemal B tubule inner sheath / axonemal A tubule inner sheath / epithelial cilium movement involved in determination of left/right asymmetry / regulation of calcineurin-NFAT signaling cascade / MAP kinase phosphatase activity / inner dynein arm assembly / Intraflagellar transport / Carboxyterminal post-translational modifications of tubulin / protein polyglutamylation / positive regulation of feeding behavior / cerebrospinal fluid circulation / sperm axoneme assembly / COPI-independent Golgi-to-ER retrograde traffic / cilium-dependent cell motility / sperm principal piece / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / regulation of cilium beat frequency involved in ciliary motility / cilium movement involved in cell motility / regulation of store-operated calcium entry / epithelial cilium movement involved in extracellular fluid movement / 9+2 motile cilium / intraciliary transport / negative regulation of chemotaxis / COPI-mediated anterograde transport / PKR-mediated signaling / Aggrephagy / 転移酵素; リンを含む基を移すもの / acrosomal membrane / Kinesins / ciliary transition zone / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / cilium movement / Resolution of Sister Chromatid Cohesion / RHO GTPases activate IQGAPs / calcium ion sensor activity / The role of GTSE1 in G2/M progression after G2 checkpoint / Recycling pathway of L1 / negative regulation of T cell activation / axoneme assembly / axonemal microtubule / left/right axis specification / cilium organization / COPI-dependent Golgi-to-ER retrograde traffic / gamma-tubulin ring complex / flagellated sperm motility / RHO GTPases Activate Formins / Separation of Sister Chromatids / Hedgehog 'off' state / organelle transport along microtubule / glial cell differentiation / Loss of Nlp from mitotic centrosomes / Recruitment of mitotic centrosome proteins and complexes / Loss of proteins required for interphase microtubule organization from the centrosome / protein tyrosine/serine/threonine phosphatase activity / Recruitment of NuMA to mitotic centrosomes / Anchoring of the basal body to the plasma membrane / AURKA Activation by TPX2 / forebrain morphogenesis / neuron projection arborization / Regulation of PLK1 Activity at G2/M Transition / positive regulation of focal adhesion disassembly / manchette / cerebellar cortex morphogenesis / positive regulation of cilium assembly / dentate gyrus development / MHC class II antigen presentation / pyramidal neuron differentiation / negative regulation of JNK cascade / CTP biosynthetic process / UTP biosynthetic process / motile cilium / positive regulation of cell motility / determination of left/right symmetry / centrosome cycle / motor behavior / GTP biosynthetic process / microtubule organizing center / response to L-glutamate / intermediate filament / regulation of neuron projection development / ciliary base / nucleoside diphosphate kinase activity / regulation of focal adhesion assembly / tubulin complex / negative regulation of T cell receptor signaling pathway / negative regulation of MAPK cascade / smoothened signaling pathway 類似検索 - 分子機能 | ||||||||||||
生物種 | ![]() ![]() | ||||||||||||
手法 | サブトモグラム平均法 / クライオ電子顕微鏡法 / 解像度: 7.7 Å | ||||||||||||
![]() | Chen Z / Shiozak M / Hass KM / Skinner W / Zhao S / Guo C / Polacco BJ / Yu Z / Krogan NJ / Kaake RM ...Chen Z / Shiozak M / Hass KM / Skinner W / Zhao S / Guo C / Polacco BJ / Yu Z / Krogan NJ / Kaake RM / Vale RD / Agard DA | ||||||||||||
資金援助 | ![]()
| ||||||||||||
![]() | ![]() タイトル: De novo protein identification in mammalian sperm using in situ cryoelectron tomography and AlphaFold2 docking. 著者: Zhen Chen / Momoko Shiozaki / Kelsey M Haas / Will M Skinner / Shumei Zhao / Caiying Guo / Benjamin J Polacco / Zhiheng Yu / Nevan J Krogan / Polina V Lishko / Robyn M Kaake / Ronald D Vale / David A Agard / ![]() 要旨: To understand the molecular mechanisms of cellular pathways, contemporary workflows typically require multiple techniques to identify proteins, track their localization, and determine their ...To understand the molecular mechanisms of cellular pathways, contemporary workflows typically require multiple techniques to identify proteins, track their localization, and determine their structures in vitro. Here, we combined cellular cryoelectron tomography (cryo-ET) and AlphaFold2 modeling to address these questions and understand how mammalian sperm are built in situ. Our cellular cryo-ET and subtomogram averaging provided 6.0-Å reconstructions of axonemal microtubule structures. The well-resolved tertiary structures allowed us to unbiasedly match sperm-specific densities with 21,615 AlphaFold2-predicted protein models of the mouse proteome. We identified Tektin 5, CCDC105, and SPACA9 as novel microtubule-associated proteins. These proteins form an extensive interaction network crosslinking the lumen of axonemal doublet microtubules, suggesting their roles in modulating the mechanical properties of the filaments. Indeed, Tekt5 -/- sperm possess more deformed flagella with 180° bends. Together, our studies presented a cellular visual proteomics workflow and shed light on the in vivo functions of Tektin 5. | ||||||||||||
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 | ![]() | 20.4 MB | ![]() | |
---|---|---|---|---|
ヘッダ (付随情報) | ![]() ![]() | 54.2 KB 54.2 KB | 表示 表示 | ![]() |
画像 | ![]() | 136 KB | ||
Filedesc metadata | ![]() | 16.2 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 566.8 KB | 表示 | ![]() |
---|---|---|---|---|
文書・詳細版 | ![]() | 566.4 KB | 表示 | |
XML形式データ | ![]() | 6.3 KB | 表示 | |
CIF形式データ | ![]() | 7.2 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 | ![]() 8to0MC C: 同じ文献を引用 ( M: このマップから作成された原子モデル |
---|---|
類似構造データ | 類似検索 - 機能・相同性 ![]() |
-
リンク
EMDBのページ | ![]() ![]() |
---|---|
「今月の分子」の関連する項目 |
-
マップ
ファイル | ![]() | ||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Final map of 48 nm-repeating unit of mouse sperm doublets | ||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 2.65 Å | ||||||||||||||||||||
密度 |
| ||||||||||||||||||||
対称性 | 空間群: 1 | ||||||||||||||||||||
詳細 | EMDB XML:
|
-添付データ
-
試料の構成要素
+全体 : Mouse sperm
+超分子 #1: Mouse sperm
+分子 #1: Cilia- and flagella-associated protein 95
+分子 #2: Tektin-1
+分子 #3: Detyrosinated tubulin alpha-1A chain
+分子 #4: Tubulin beta-4B chain
+分子 #5: Tektin-2
+分子 #6: EF-hand domain-containing family member B
+分子 #7: Tektin-3
+分子 #8: Meiosis-specific nuclear structural protein 1
+分子 #9: Tektin-4
+分子 #10: Cilia- and flagella-associated protein 53
+分子 #11: Tektin bundle-interacting protein 1
+分子 #12: Cilia- and flagella-associated protein 141
+分子 #13: Nucleoside diphosphate kinase 7
+分子 #14: Protein FAM166B
+分子 #15: Sperm-associated antigen 8
+分子 #16: RIB43A-like with coiled-coils protein 2
+分子 #17: Cilia- and flagella-associated protein 107
+分子 #18: Cilia- and flagella-associated protein 161
+分子 #19: EF-hand domain-containing protein 1
+分子 #20: Sperm acrosome-associated protein 9
+分子 #21: EF-hand domain-containing family member C2
+分子 #22: Cilia- and flagella-associated protein 20
+分子 #23: Parkin coregulated gene protein homolog
+分子 #24: Dual specificity protein phosphatase 3
+分子 #25: Piercer of microtubule wall 2 protein
+分子 #26: Tektin-5
+分子 #27: Cilia- and flagella-associated protein 45
+分子 #28: Cilia- and flagella-associated protein 52
+分子 #29: Enkurin
+分子 #30: Protein Flattop
+分子 #31: Cilia- and flagella- associated protein 210
+分子 #32: Cilia- and flagella-associated protein 276
+分子 #33: EF-hand calcium-binding domain-containing protein 6
+分子 #34: Coiled-coil domain-containing protein 105
+分子 #35: Piercer of microtubule wall 1 protein
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
---|---|
![]() | サブトモグラム平均法 |
試料の集合状態 | cell |
-
試料調製
緩衝液 | pH: 7.4 |
---|---|
凍結 | 凍結剤: ETHANE |
-
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: 4.0 e/Å2 |
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 / 最大 デフォーカス(公称値): 6.0 µm / 最小 デフォーカス(公称値): 2.0 µm |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
-
画像解析
最終 再構成 | 想定した対称性 - 点群: C1 (非対称) / 解像度のタイプ: BY AUTHOR / 解像度: 7.7 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: RELION (ver. 4.0-beta2) / 使用したサブトモグラム数: 12848 |
---|---|
抽出 | トモグラム数: 76 / 使用した粒子像数: 32288 詳細: 32288 particles were initially picked every 24 nm along the microtubules. |
最終 角度割当 | タイプ: MAXIMUM LIKELIHOOD |