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TitleHigh-resolution cryo-EM analysis of the yeast ATP synthase in a lipid membrane.
Journal, issue, pagesScience, Vol. 360, Issue 6389, Year 2018
Publish dateMay 11, 2018
AuthorsAnurag P Srivastava / Min Luo / Wenchang Zhou / Jindrich Symersky / Dongyang Bai / Melissa G Chambers / José D Faraldo-Gómez / Maofu Liao / David M Mueller /
PubMed AbstractMitochondrial adenosine triphosphate (ATP) synthase comprises a membrane embedded F motor that rotates to drive ATP synthesis in the F subunit. We used single-particle cryo-electron microscopy (cryo- ...Mitochondrial adenosine triphosphate (ATP) synthase comprises a membrane embedded F motor that rotates to drive ATP synthesis in the F subunit. We used single-particle cryo-electron microscopy (cryo-EM) to obtain structures of the full complex in a lipid bilayer in the absence or presence of the inhibitor oligomycin at 3.6- and 3.8-angstrom resolution, respectively. To limit conformational heterogeneity, we locked the rotor in a single conformation by fusing the F6 subunit of the stator with the δ subunit of the rotor. Assembly of the enzyme with the F6-δ fusion caused a twisting of the rotor and a 9° rotation of the F c-ring in the direction of ATP synthesis, relative to the structure of isolated F Our cryo-EM structures show how F and F are coupled, give insight into the proton translocation pathway, and show how oligomycin blocks ATP synthesis.
External linksScience / PubMed:29650704 / PubMed Central
MethodsEM (single particle)
Resolution3.6 - 4.2 Å
Structure data

7546

6cp3

EMDB-7546, PDB-6cp3:
Monomer yeast ATP synthase (F1Fo) reconstituted in nanodisc with inhibitor of oligomycin bound.
Method: EM (single particle) / Resolution: 3.8 Å

7547

6cp5

EMDB-7547, PDB-6cp5:
Monomer yeast ATP synthase Fo reconstituted in nanodisc with inhibitor of oligomycin bound generated from focused refinement.
Method: EM (single particle) / Resolution: 4.2 Å

7548

6cp6

EMDB-7548, PDB-6cp6:
Monomer yeast ATP synthase (F1Fo) reconstituted in nanodisc.
Method: EM (single particle) / Resolution: 3.6 Å

7549

6cp7

EMDB-7549, PDB-6cp7:
Monomer yeast ATP synthase Fo reconstituted in nanodisc generated from masked refinement.
Method: EM (single particle) / Resolution: 4.1 Å

Chemicals

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

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

ChemComp-EFO:
Oligomycin A

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
KeywordsBIOSYNTHETIC PROTEIN / ATP synthase

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