+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 7nie | ||||||
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タイトル | putative glycerol kinase-like proteins anchored on an array of voltage dependent anion channels in the outer mitochondrial membrane of pig sperm mitochondria | ||||||
要素 |
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キーワード | MEMBRANE PROTEIN / mitochondria / sperm | ||||||
機能・相同性 | 機能・相同性情報 PINK1-PRKN Mediated Mitophagy / Triglyceride biosynthesis / Ub-specific processing proteases / glycerol-3-phosphate biosynthetic process / voltage-gated monoatomic anion channel activity / mitochondrial outer membrane permeabilization / glycerol kinase / glycerol kinase activity / glycerol metabolic process / ceramide binding ...PINK1-PRKN Mediated Mitophagy / Triglyceride biosynthesis / Ub-specific processing proteases / glycerol-3-phosphate biosynthetic process / voltage-gated monoatomic anion channel activity / mitochondrial outer membrane permeabilization / glycerol kinase / glycerol kinase activity / glycerol metabolic process / ceramide binding / phosphatidylcholine binding / oxysterol binding / glycerol catabolic process / triglyceride metabolic process / lipid transport / porin activity / cholesterol binding / pore complex / sperm midpiece / mitochondrial outer membrane / mitochondrion / ATP binding / membrane 類似検索 - 分子機能 | ||||||
生物種 | Sus scrofa (ブタ) | ||||||
手法 | 電子顕微鏡法 / サブトモグラム平均法 / クライオ電子顕微鏡法 / 解像度: 35 Å | ||||||
データ登録者 | Leung, M.R. / Zeev-Ben-Mordehai, T. | ||||||
資金援助 | オランダ, 1件
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引用 | ジャーナル: Proc Natl Acad Sci U S A / 年: 2021 タイトル: In-cell structures of conserved supramolecular protein arrays at the mitochondria-cytoskeleton interface in mammalian sperm. 著者: Miguel Ricardo Leung / Riccardo Zenezini Chiozzi / Marc C Roelofs / Johannes F Hevler / Ravi Teja Ravi / Paula Maitan / Min Zhang / Heiko Henning / Elizabeth G Bromfield / Stuart C Howes / ...著者: Miguel Ricardo Leung / Riccardo Zenezini Chiozzi / Marc C Roelofs / Johannes F Hevler / Ravi Teja Ravi / Paula Maitan / Min Zhang / Heiko Henning / Elizabeth G Bromfield / Stuart C Howes / Bart M Gadella / Albert J R Heck / Tzviya Zeev-Ben-Mordehai / 要旨: Mitochondria-cytoskeleton interactions modulate cellular physiology by regulating mitochondrial transport, positioning, and immobilization. However, there is very little structural information ...Mitochondria-cytoskeleton interactions modulate cellular physiology by regulating mitochondrial transport, positioning, and immobilization. However, there is very little structural information defining mitochondria-cytoskeleton interfaces in any cell type. Here, we use cryofocused ion beam milling-enabled cryoelectron tomography to image mammalian sperm, where mitochondria wrap around the flagellar cytoskeleton. We find that mitochondria are tethered to their neighbors through intermitochondrial linkers and are anchored to the cytoskeleton through ordered arrays on the outer mitochondrial membrane. We use subtomogram averaging to resolve in-cell structures of these arrays from three mammalian species, revealing they are conserved across species despite variations in mitochondrial dimensions and cristae organization. We find that the arrays consist of boat-shaped particles anchored on a network of membrane pores whose arrangement and dimensions are consistent with voltage-dependent anion channels. Proteomics and in-cell cross-linking mass spectrometry suggest that the conserved arrays are composed of glycerol kinase-like proteins. Ordered supramolecular assemblies may serve to stabilize similar contact sites in other cell types in which mitochondria need to be immobilized in specific subcellular environments, such as in muscles and neurons. | ||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | 分子: MolmilJmol/JSmol |
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 7nie.cif.gz | 1.2 MB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb7nie.ent.gz | 1013.7 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 7nie.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
その他 | その他のダウンロード |
-検証レポート
文書・要旨 | 7nie_validation.pdf.gz | 798.7 KB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 7nie_full_validation.pdf.gz | 854.7 KB | 表示 | |
XML形式データ | 7nie_validation.xml.gz | 130 KB | 表示 | |
CIF形式データ | 7nie_validation.cif.gz | 193.5 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/ni/7nie ftp://data.pdbj.org/pub/pdb/validation_reports/ni/7nie | HTTPS FTP |
-関連構造データ
-リンク
-集合体
登録構造単位 |
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1 |
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-要素
#1: タンパク質 | 分子量: 53579.547 Da / 分子数: 4 / 由来タイプ: 天然 / 由来: (天然) Sus scrofa (ブタ) Plasmid details: lamellae prepared by cryo-focused ion beam milling of pig sperm 組織: sperm / 参照: UniProt: A0A287BD08, glycerol kinase #2: タンパク質 | 分子量: 31637.484 Da / 分子数: 12 / 由来タイプ: 天然 / 由来: (天然) Sus scrofa (ブタ) Plasmid details: lamellae prepared by cryo-focused ion beam milling of pig sperm 組織: sperm / 参照: UniProt: F1S2F6 Has protein modification | Y | |
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-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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EM実験 | 試料の集合状態: CELL / 3次元再構成法: サブトモグラム平均法 |
-試料調製
構成要素 | 名称: subtomogram average of ordered protein arrays on the axoneme-facing surface of the outer mitochondrial membrane in pig sperm mitochondria (alignment focused on one unit) タイプ: CELL / Entity ID: all / 由来: NATURAL |
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由来(天然) | 生物種: Sus scrofa (ブタ) / 細胞内の位置: midpiece / Organelle: mitochondria / 組織: sperm |
緩衝液 | pH: 7.4 |
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
試料支持 | グリッドの材料: COPPER / グリッドのサイズ: 200 divisions/in. / グリッドのタイプ: Quantifoil |
急速凍結 | 装置: HOMEMADE PLUNGER / 凍結剤: ETHANE-PROPANE |
-電子顕微鏡撮影
実験機器 | モデル: Talos Arctica / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TALOS ARCTICA |
電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 200 kV / 照射モード: FLOOD BEAM |
電子レンズ | モード: BRIGHT FIELD / アライメント法: COMA FREE |
試料ホルダ | 凍結剤: NITROGEN |
撮影 | 電子線照射量: 1.3 e/Å2 / 検出モード: COUNTING フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) |
電子光学装置 | エネルギーフィルタースリット幅: 20 eV |
-解析
EMソフトウェア |
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CTF補正 | タイプ: PHASE FLIPPING ONLY | ||||||||||||||||||||||||||||
対称性 | 点対称性: C2 (2回回転対称) | ||||||||||||||||||||||||||||
3次元再構成 | 解像度: 35 Å / 解像度の算出法: FSC 0.5 CUT-OFF / 粒子像の数: 536 / クラス平均像の数: 1 / 対称性のタイプ: POINT | ||||||||||||||||||||||||||||
EM volume selection | Num. of tomograms: 3 / Num. of volumes extracted: 268 / Reference model: random particle from the dataset | ||||||||||||||||||||||||||||
原子モデル構築 | プロトコル: RIGID BODY FIT 詳細: Homology models of pig glycerol kinase and VDAC2 were made using Robetta. Rigid body fitting was done with Chimera/ChimeraX. Putative GK proteins only fit in one orientation, but for VDAC ...詳細: Homology models of pig glycerol kinase and VDAC2 were made using Robetta. Rigid body fitting was done with Chimera/ChimeraX. Putative GK proteins only fit in one orientation, but for VDAC proteins the rotation around the pore axis remains ambiguous. |