7U8P

Structure of porcine kidney V-ATPase with SidK, Rotary State 1

This is a non-PDB format compatible entry.

Summary for 7U8P

• Entry DOI: 10.2210/pdb7u8p/pdb
• EMDB information: 26386
• Descriptor: V-type proton ATPase catalytic subunit A, V-type proton ATPase subunit a, V-type proton ATPase 21 kDa proteolipid subunit isoform 1, ... (18 entities in total)
• Functional Keywords: proton translocation, complex, membrane protein
• Biological source: Legionella pneumophila; More
• Total number of polymer chains: 35
• Total formula weight: 1164267.11

Authors

Tan, Y.Z.,Keon, K.A. (deposition date: 2022-03-09, release date: 2022-07-06, Last modification date: 2024-02-14)

Primary citation

Tan, Y.Z.,Abbas, Y.M.,Wu, J.Z.,Wu, D.,Keon, K.A.,Hesketh, G.G.,Bueler, S.A.,Gingras, A.C.,Robinson, C.V.,Grinstein, S.,Rubinstein, J.L.
CryoEM of endogenous mammalian V-ATPase interacting with the TLDc protein mEAK-7.
Life Sci Alliance, 5:-, 2022

PubMed Abstract: 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.

PubMed: 35794005
DOI: 10.26508/lsa.202201527

Experimental method

ELECTRON MICROSCOPY (3.7 Å)