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-Structure paper
タイトル | CryoEM of endogenous mammalian V-ATPase interacting with the TLDc protein mEAK-7. |
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ジャーナル・号・ページ | Life Sci Alliance, Vol. 5, Issue 11, Year 2022 |
掲載日 | 2022年7月6日 |
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 / |
PubMed 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. |
リンク | Life Sci Alliance / PubMed:35794005 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 3.5 - 4.1 Å |
構造データ | EMDB-26385, PDB-7u8o: EMDB-26386, PDB-7u8p: EMDB-26387, PDB-7u8q: EMDB-26388, PDB-7u8r: |
化合物 | ChemComp-ADP: |
由来 |
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キーワード | MEMBRANE PROTEIN / proton translocation / complex / TLDc |