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TitleStructure of V-ATPase from the mammalian brain.
Journal, issue, pagesScience, Vol. 367, Issue 6483, Page 1240-1246, Year 2020
Publish dateMar 13, 2020
AuthorsYazan M Abbas / Di Wu / Stephanie A Bueler / Carol V Robinson / John L Rubinstein /
PubMed AbstractIn neurons, the loading of neurotransmitters into synaptic vesicles uses energy from proton-pumping vesicular- or vacuolar-type adenosine triphosphatases (V-ATPases). These membrane protein complexes ...In neurons, the loading of neurotransmitters into synaptic vesicles uses energy from proton-pumping vesicular- or vacuolar-type adenosine triphosphatases (V-ATPases). These membrane protein complexes possess numerous subunit isoforms, which complicates their analysis. We isolated homogeneous rat brain V-ATPase through its interaction with SidK, a effector protein. Cryo-electron microscopy allowed the construction of an atomic model, defining the enzyme's ATP:proton ratio as 3:10 and revealing a homolog of yeast subunit f in the membrane region, which we tentatively identify as RNAseK. The c ring encloses the transmembrane anchors for cleaved ATP6AP1/Ac45 and ATP6AP2/PRR, the latter of which is the (pro)renin receptor that, in other contexts, is involved in both Wnt signaling and the renin-angiotensin system that regulates blood pressure. This structure shows how ATP6AP1/Ac45 and ATP6AP2/PRR enable assembly of the enzyme's catalytic and membrane regions.
External linksScience / PubMed:32165585 / PubMed Central
MethodsEM (single particle)
Resolution3.6 - 5.7 Å
Structure data

EMDB-21317: Mammalian V-ATPase from rat brain with the Legionella pneumophila effector protein SidK - rotational state 1 non-uniform refinement
PDB-6vq6: Mammalian V-ATPase from rat brain - composite model of rotational state 1 bound to ADP and SidK (built from focused refinement models)
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-21318: Mammalian V-ATPase from rat brain with the Legionella pneumophila effector protein SidK - rotational state 2 non-uniform refinement
PDB-6vq7: Mammalian V-ATPase from rat brain - composite model of rotational state 2 bound to ADP and SidK (built from focused refinement models)
Method: EM (single particle) / Resolution: 4.0 Å

EMDB-21319: Mammalian V-ATPase from rat brain with the Legionella pneumophila effector protein SidK - rotational state 3 non-uniform refinement
PDB-6vq8: Mammalian V-ATPase from rat brain - composite model of rotational state 3 bound to ADP and SidK (built from focused refinement models)
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-21345, PDB-6vq9:
Mammalian V-ATPase from rat brain soluble V1 region rotational state 1 with SidK and ADP (from focused refinement)
Method: EM (single particle) / Resolution: 3.6 Å

EMDB-21346, PDB-6vqa:
Mammalian V-ATPase from rat brain soluble V1 region rotational state 2 with SidK and ADP (from focused refinement)
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-21347, PDB-6vqb:
Mammalian V-ATPase from rat brain soluble V1 region rotational state 2 with SidK and ADP (from focused refinement)
Method: EM (single particle) / Resolution: 3.6 Å

EMDB-21348, PDB-6vqc:
Mammalian V-ATPase from rat brain membrane-embedded Vo region rotational state 1 (from focused refinement)
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-21349, PDB-6vqg:
Mammalian V-ATPase from rat brain membrane-embedded Vo region rotational state 2 (from focused refinement)
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-21350, PDB-6vqh:
Mammalian V-ATPase from rat brain membrane-embedded Vo region rotational state 3 (from focused refinement)
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-21351, PDB-6vqi:
Mammalian V-ATPase from rat brain collar and peripheral stalks rotational state 1 (from focused refinement)
Method: EM (single particle) / Resolution: 4.3 Å

EMDB-21352, PDB-6vqj:
Mammalian V-ATPase from rat brain collar and peripheral stalks rotational state 2 (from focused refinement)
Method: EM (single particle) / Resolution: 5.7 Å

EMDB-21353, PDB-6vqk:
Mammalian V-ATPase from rat brain collar and peripheral stalks rotational state 3 (from focused refinement)
Method: EM (single particle) / Resolution: 5.7 Å

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

ChemComp-MG:
Unknown entry

Source
  • rattus norvegicus (Norway rat)
  • legionella pneumophila subsp. pneumophila (strain philadelphia 1 / atcc 33152 / dsm 7513) (bacteria)
KeywordsPROTON TRANSPORT / membrane protein complex / rotary atpase

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