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データを開く
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基本情報
登録情報 | ![]() | |||||||||||||||
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タイトル | Structure of porcine kidney V-ATPase with SidK, Rotary State 2 | |||||||||||||||
![]() | Map used for model building. | |||||||||||||||
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![]() | proton translocation / ![]() ![]() | |||||||||||||||
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 / transporter activator activity / Insulin receptor recycling / cellular response to increased oxygen levels / proton-transporting V-type ATPase, V1 domain / synaptic vesicle lumen acidification ...ROS and RNS production in phagocytes / RHOA GTPase cycle / Transferrin endocytosis and recycling / Amino acids regulate mTORC1 / Ion channel transport / transporter activator activity / Insulin receptor recycling / cellular response to increased oxygen levels / proton-transporting V-type ATPase, V1 domain / synaptic vesicle lumen acidification / endosome to plasma membrane protein transport / proton-transporting V-type ATPase, V0 domain / plasma membrane proton-transporting V-type ATPase complex / proton-transporting two-sector ATPase complex, catalytic domain / 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 / vacuolar proton-transporting V-type ATPase complex / cell projection organization / vacuolar acidification / dendritic spine membrane / regulation of cellular pH / vacuolar membrane / osteoclast development / transmembrane transporter complex / ATPase activator activity / autophagosome membrane / ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() 類似検索 - 分子機能 | |||||||||||||||
生物種 | ![]() ![]() ![]() ![]() ![]() ![]() | |||||||||||||||
手法 | ![]() ![]() | |||||||||||||||
![]() | Tan YZ | |||||||||||||||
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
添付画像 |
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ダウンロードとリンク
-EMDBアーカイブ
マップデータ | ![]() | 12 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 35.6 KB 35.6 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 10.4 KB | 表示 | ![]() |
画像 | ![]() | 56 KB | ||
Filedesc metadata | ![]() | 9.9 KB | ||
その他 | ![]() ![]() ![]() | 97.4 MB 95.5 MB 95.5 MB | ||
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 | ![]() 7u8qMC ![]() 7u8oC ![]() 7u8pC ![]() 7u8rC M: このマップから作成された原子モデル 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
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 | Map used for model building. | ||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.52801 Å | ||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-追加マップ: V-ATPase with SidK, Rotary State 2
ファイル | emd_26387_additional_1.map | ||||||||||||
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注釈 | V-ATPase with SidK, Rotary State 2 | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: V-ATPase with SidK, Rotary State 2
ファイル | emd_26387_half_map_1.map | ||||||||||||
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注釈 | V-ATPase with SidK, Rotary State 2 | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: V-ATPase with SidK, Rotary State 2
ファイル | emd_26387_half_map_2.map | ||||||||||||
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注釈 | V-ATPase with SidK, Rotary State 2 | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
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試料の構成要素
+全体 : Porcine kidney V-ATPase with SidK, Rotary State 2
+超分子 #1: Porcine kidney V-ATPase with SidK, Rotary State 2
+分子 #1: V-type proton ATPase catalytic subunit A
+分子 #2: Vacuolar proton pump subunit B
+分子 #3: V-type proton ATPase subunit C
+分子 #4: V-type proton ATPase subunit D
+分子 #5: V-type proton ATPase subunit E 1
+分子 #6: V-type proton ATPase subunit F
+分子 #7: V-type proton ATPase subunit G
+分子 #8: Bacterial effector protein SidK
+分子 #9: V-type proton ATPase subunit H
+分子 #10: V-type proton ATPase subunit a
+分子 #11: V-type proton ATPase 21 kDa proteolipid subunit isoform 1
+分子 #12: ATPase H+ transporting accessory protein 1
+分子 #13: V-type proton ATPase subunit
+分子 #14: V-type proton ATPase subunit
+分子 #15: Ribonuclease kappa
+分子 #16: V-type proton ATPase proteolipid subunit
+分子 #17: ATPase H(+)-transporting lysosomal accessory protein 2
+分子 #18: ADENOSINE-5'-DIPHOSPHATE
-実験情報
-構造解析
手法 | ![]() |
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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 |
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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 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD![]() |
撮影 | フィルム・検出器のモデル: FEI FALCON IV (4k x 4k) 平均電子線量: 40.0 e/Å2 |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |