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TitleOxidative stress protein Oxr1 promotes V-ATPase holoenzyme disassembly in catalytic activity-independent manner.
Journal, issue, pagesEMBO J, Vol. 41, Issue 3, Page e109360, Year 2022
Publish dateFeb 1, 2022
AuthorsMd Murad Khan / Seowon Lee / Sergio Couoh-Cardel / Rebecca A Oot / Hyunmin Kim / Stephan Wilkens / Soung-Hun Roh /
PubMed AbstractThe vacuolar ATPase (V-ATPase) is a rotary motor proton pump that is regulated by an assembly equilibrium between active holoenzyme and autoinhibited V -ATPase and V proton channel subcomplexes. ...The vacuolar ATPase (V-ATPase) is a rotary motor proton pump that is regulated by an assembly equilibrium between active holoenzyme and autoinhibited V -ATPase and V proton channel subcomplexes. Here, we report cryo-EM structures of yeast V-ATPase assembled in vitro from lipid nanodisc reconstituted V and mutant V . Our analysis identified holoenzymes in three active rotary states, indicating that binding of V to V provides sufficient free energy to overcome V autoinhibition. Moreover, the structures suggest that the unequal spacing of V 's proton-carrying glutamic acid residues serves to alleviate the symmetry mismatch between V and V motors, a notion that is supported by mutagenesis experiments. We also uncover a structure of free V bound to Oxr1, a conserved but poorly characterized factor involved in the oxidative stress response. Biochemical experiments show that Oxr1 inhibits V -ATPase and causes disassembly of the holoenzyme, suggesting that Oxr1 plays a direct role in V-ATPase regulation.
External linksEMBO J / PubMed:34918374 / PubMed Central
MethodsEM (single particle)
Resolution3.8 - 6.6 Å
Structure data

31538

7fda

EMDB-31538, PDB-7fda:
CryoEM Structure of Reconstituted V-ATPase, state1
Method: EM (single particle) / Resolution: 4.2 Å

31539

7fdb

EMDB-31539, PDB-7fdb:
CryoEM Structures of Reconstituted V-ATPase,State2
Method: EM (single particle) / Resolution: 4.8 Å

31540

7fdc

EMDB-31540, PDB-7fdc:
CryoEM Structures of Reconstituted V-ATPase, state3
Method: EM (single particle) / Resolution: 6.6 Å

31541

7fde

EMDB-31541, PDB-7fde:
CryoEM Structures of Reconstituted V-ATPase, Oxr1 bound V1
Method: EM (single particle) / Resolution: 3.8 Å

Source
  • saccharomyces cerevisiae s288c (yeast)
  • homo sapiens (human)
KeywordsMOTOR PROTEIN / ATPase / proton pump / rotary motor enzyme / membrane protein

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