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基本情報
登録情報 | データベース: PDB / ID: 7u8o | |||||||||||||||
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タイトル | Structure of porcine V-ATPase with mEAK7 and SidK, Rotary state 2 | |||||||||||||||
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![]() | MEMBRANE PROTEIN / proton translocation / complex / TLDc | |||||||||||||||
機能・相同性 | ![]() ROS and RNS production in phagocytes / RHOA GTPase cycle / Transferrin endocytosis and recycling / Amino acids regulate mTORC1 / Ion channel transport / Insulin receptor recycling / transporter activator activity / plasma membrane proton-transporting V-type ATPase complex / cellular response to increased oxygen levels / proton-transporting V-type ATPase, V1 domain ...ROS and RNS production in phagocytes / RHOA GTPase cycle / Transferrin endocytosis and recycling / Amino acids regulate mTORC1 / Ion channel transport / Insulin receptor recycling / transporter activator activity / plasma membrane proton-transporting V-type ATPase complex / cellular response to increased oxygen levels / proton-transporting V-type ATPase, V1 domain / synaptic vesicle lumen acidification / proton-transporting V-type ATPase, V0 domain / proton-transporting two-sector ATPase complex, catalytic domain / endosome to plasma membrane protein transport / clathrin-coated vesicle membrane / vacuolar proton-transporting V-type ATPase, V0 domain / vacuolar proton-transporting V-type ATPase, V1 domain / vacuolar transport / proton-transporting V-type ATPase complex / cell projection organization / osteoclast development / vacuolar proton-transporting V-type ATPase complex / dendritic spine membrane / regulation of cellular pH / vacuolar acidification / vacuolar membrane / microvillus / autophagosome membrane / ATPase activator activity / transmembrane transporter complex / ATP metabolic process / H+-transporting two-sector ATPase / transport vesicle / RNA endonuclease activity / proton-transporting ATPase activity, rotational mechanism / proton transmembrane transport / proton-transporting ATP synthase activity, rotational mechanism / small GTPase binding / synaptic vesicle membrane / melanosome / presynapse / signaling receptor activity / ATPase binding / intracellular iron ion homeostasis / early endosome / lysosome / endosome membrane / endosome / apical plasma membrane / lysosomal membrane / endoplasmic reticulum membrane / ATP hydrolysis activity / ATP binding / membrane / plasma membrane / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||||||||
生物種 | ![]() ![]() ![]() ![]() | |||||||||||||||
手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.5 Å | |||||||||||||||
![]() | Tan, Y.Z. / Keon, K.A. | |||||||||||||||
資金援助 | ![]() ![]()
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![]() | ![]() タイトル: CryoEM of endogenous mammalian V-ATPase interacting with the TLDc protein mEAK-7. 著者: Yong Zi Tan / Yazan M Abbas / Jing Ze Wu / Di Wu / Kristine A Keon / Geoffrey G Hesketh / Stephanie A Bueler / Anne-Claude Gingras / Carol V Robinson / Sergio Grinstein / John L Rubinstein / ![]() ![]() 要旨: V-ATPases are rotary proton pumps that serve as signaling hubs with numerous protein binding partners. CryoEM with exhaustive focused classification allowed detection of endogenous proteins ...V-ATPases are rotary proton pumps that serve as signaling hubs with numerous protein binding partners. CryoEM with exhaustive focused classification allowed detection of endogenous proteins associated with porcine kidney V-ATPase. An extra C subunit was found in ∼3% of complexes, whereas ∼1.6% of complexes bound mEAK-7, a protein with proposed roles in dauer formation in nematodes and mTOR signaling in mammals. High-resolution cryoEM of porcine kidney V-ATPase with recombinant mEAK-7 showed that mEAK-7's TLDc domain interacts with V-ATPase's stator, whereas its C-terminal α helix binds V-ATPase's rotor. This crosslink would be expected to inhibit rotary catalysis. However, unlike the yeast TLDc protein Oxr1p, exogenous mEAK-7 does not inhibit V-ATPase and mEAK-7 overexpression in cells does not alter lysosomal or phagosomal pH. Instead, cryoEM suggests that the mEAK-7:V-ATPase interaction is disrupted by ATP-induced rotation of the rotor. Comparison of Oxr1p and mEAK-7 binding explains this difference. These results show that V-ATPase binding by TLDc domain proteins can lead to effects ranging from strong inhibition to formation of labile interactions that are sensitive to the enzyme's activity. | |||||||||||||||
履歴 |
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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ダウンロードとリンク
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ダウンロード
PDBx/mmCIF形式 | ![]() | 1.5 MB | 表示 | ![]() |
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PDB形式 | ![]() | 表示 | ![]() | |
PDBx/mmJSON形式 | ![]() | ツリー表示 | ![]() | |
その他 | ![]() |
-検証レポート
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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-関連構造データ
関連構造データ | ![]() 26385MC ![]() 7u8pC ![]() 7u8qC ![]() 7u8rC M: このデータのモデリングに利用したマップデータ C: 同じ文献を引用 ( |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
電子顕微鏡画像生データ | ![]() Data size: 12.7 TB Data #1: Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7 collected using Tundra [micrographs - multiframe] Data #2: Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7 collected using Tundra [micrographs - single frame] Data #3: Polished particles of Pig Kidney V-ATPase bound to mEAK-7 collected using Tundra [picked particles - single frame - processed] Data #4: Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7 collected using Titan Krios and Falcon4 [micrographs - multiframe] Data #5: Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7 collected using Titan Krios and Falcon4 [micrographs - single frame] Data #6: Polished particles of Pig Kidney V-ATPase bound to mEAK-7 collected using Titan Krios and Falcon4 [picked particles - multiframe - processed] Data #7: Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7deltaCterm collected using Titan Krios and Falcon4 [micrographs - multiframe] Data #8: Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7deltaCterm collected using Titan Krios and Falcon4 [micrographs - single frame] Data #9: Polished particles of Pig Kidney V-ATPase bound to mEAK-7deltaCterm collected using Titan Krios and Falcon4 [picked particles - single frame - processed] Data #10: Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7 with ATP collected using Titan Krios and Falcon4 [micrographs - multiframe] Data #11: Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7 with ATP collected using Titan Krios and Falcon4 [micrographs - single frame] Data #12: Polished particles of Pig Kidney V-ATPase bound to mEAK-7 with ATP collected using Titan Krios and Falcon4 [picked particles - single frame - processed] Data #13: Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7 with EDTA/EGTA collected using Titan Krios and Falcon4 [micrographs - multiframe] Data #14: Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7 with EDTA/EGTA collected using Titan Krios and Falcon4 [micrographs - single frame] Data #15: Polished particles of Pig Kidney V-ATPase bound to mEAK-7 with EDTA/EGTA collected using Titan Krios and Falcon4 [picked particles - single frame - processed] Data #16: Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7 with Calcium collected using Glacios with Selectris X and Falcon 4 [micrographs - multiframe] Data #17: Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7 with Calcium collected using Glacios with Selectris X and Falcon 4 [micrographs - single frame] Data #18: Polished particles of Pig Kidney V-ATPase bound to mEAK-7 with Calcium collected using Glacios with Selectris X and Falcon 4 [picked particles - single frame - processed]) |
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リンク
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集合体
登録構造単位 | ![]()
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要素
-V-type proton ATPase ... , 12種, 26分子 ABCGHIJKLMNOTabdeghijklmno
#1: タンパク質 | 分子量: 68393.844 Da / 分子数: 3 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #3: タンパク質 | | 分子量: 44066.566 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #4: タンパク質 | | 分子量: 28301.902 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #5: タンパク質 | 分子量: 26162.373 Da / 分子数: 3 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #6: タンパク質 | | 分子量: 13403.288 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #7: タンパク質 | 分子量: 13748.474 Da / 分子数: 3 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #9: タンパク質 | | 分子量: 55917.797 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #11: タンパク質 | | 分子量: 96365.258 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #12: タンパク質 | | 分子量: 21530.426 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #14: タンパク質 | | 分子量: 40369.949 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #15: タンパク質 | | 分子量: 9343.286 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #17: タンパク質 | 分子量: 15639.677 Da / 分子数: 9 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() |
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-タンパク質 , 6種, 10分子 DEFQRSUcfp
#2: タンパク質 | 分子量: 57162.859 Da / 分子数: 3 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #8: タンパク質 | 分子量: 38539.371 Da / 分子数: 3 / 由来タイプ: 組換発現 由来: (組換発現) ![]() ![]() 発現宿主: ![]() ![]() #10: タンパク質 | | 分子量: 50380.816 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) ![]() ![]() ![]() ![]() #13: タンパク質 | | 分子量: 51547.465 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #16: タンパク質 | | 分子量: 11016.065 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #18: タンパク質 | | 分子量: 39200.055 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() |
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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: PARTICLE / 3次元再構成法: 単粒子再構成法 |
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試料調製
構成要素 | 名称: V-ATPase in complex with recombinant SidK and recombinant mEAK-7 タイプ: COMPLEX / Entity ID: all / 由来: MULTIPLE SOURCES |
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由来(天然) | 生物種: ![]() ![]() |
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 |
急速凍結 | 凍結剤: ETHANE |
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電子顕微鏡撮影
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TITAN KRIOS |
電子銃 | 電子線源: ![]() |
電子レンズ | モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2554.877 nm / 最小 デフォーカス(公称値): 787.921 nm |
撮影 | 電子線照射量: 36 e/Å2 フィルム・検出器のモデル: FEI FALCON IV (4k x 4k) |
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解析
CTF補正 | タイプ: NONE |
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3次元再構成 | 解像度: 3.5 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 31814 / 対称性のタイプ: POINT |