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Yorodumi- EMDB-74150: CryoEM structure of respiratory complex III intermediate from Aca... -
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Basic information
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| Title | CryoEM structure of respiratory complex III intermediate from Acanthamoeba castellanii | |||||||||
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Keywords | complex III / OXPHOS / cytochrome bc1 complex / OXIDOREDUCTASE | |||||||||
| Function / homology | Function and homology informationpitrilysin / signal peptidase activity / respiratory chain complex III / quinol-cytochrome-c reductase activity / mitochondrial electron transport, ubiquinol to cytochrome c / metalloendopeptidase activity / mitochondrial inner membrane / mitochondrial matrix / metal ion binding / mitochondrion ...pitrilysin / signal peptidase activity / respiratory chain complex III / quinol-cytochrome-c reductase activity / mitochondrial electron transport, ubiquinol to cytochrome c / metalloendopeptidase activity / mitochondrial inner membrane / mitochondrial matrix / metal ion binding / mitochondrion / proteolysis / nucleus Similarity search - Function | |||||||||
| Biological species | Acanthamoeba castellanii (eukaryote) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.65 Å | |||||||||
Authors | Fry MY / Luce BE / Stefely JA / Samuelson J / Mootha VK / Chao LH | |||||||||
| Funding support | United States, 1 items
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Citation | Journal: Cell / Year: 2026Title: Acanthamoeba ATP synthase structure reveals the TCA cycle is tethered to OXPHOS. Authors: Michelle Y Fry / Bridget E Luce / Jonathan A Stefely / Michael Z Chen / Felicia G Deng / Tiana M Issa / / Sarah E Calvo / John Samuelson / Vamsi K Mootha / Luke H Chao / ![]() Abstract: Oxidative phosphorylation (OXPHOS) is a key metabolic process that couples redox energy to ATP production. While some core OXPHOS complex subunits are found across all domains of life, many have ...Oxidative phosphorylation (OXPHOS) is a key metabolic process that couples redox energy to ATP production. While some core OXPHOS complex subunits are found across all domains of life, many have diverged or expanded across evolution-as seen in the protozoan pathogen Acanthamoeba castellanii. By integrating cryo-electron microscopy of unenriched mitochondrial lysate with mass spectrometry proteomics, we resolved the structures of endogenous mitochondrial ATP synthase (complex V), Hsp60, and respiratory complex III from Acanthamoeba. We capture Acanthamoeba ATP synthase in an IF1-inhibited state and reveal how Acanthamoeba-specific subunits and extensions stabilize the molecular machine, which includes a β subunit extension that interfaces with the peripheral stalk. Additionally, we characterize an active malate dehydrogenase (MDH) dimer structurally integrated within the ATP synthase peripheral stalk, thus revealing a direct protein tether between OXPHOS and the tricarboxylic acid cycle. Together, these findings provide structural insight into lineage-specific adaptations in Acanthamoeba that may tune protozoan metabolism. | |||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_74150.map.gz | 230 MB | EMDB map data format | |
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| Header (meta data) | emd-74150-v30.xml emd-74150.xml | 27.3 KB 27.3 KB | Display Display | EMDB header |
| Images | emd_74150.png | 57.9 KB | ||
| Filedesc metadata | emd-74150.cif.gz | 7.6 KB | ||
| Others | emd_74150_half_map_1.map.gz emd_74150_half_map_2.map.gz | 226.6 MB 226.6 MB | ||
| Archive directory | https://data.pdbj.org/pub/emdb/structures/EMD-74150 ftp://data.pdbj.org/pub/emdb/structures/EMD-74150 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9zfsMC ![]() 9zfrC ![]() 9zftC ![]() 9zfuC ![]() 9zfvC ![]() 9zfwC ![]() 9zfxC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_74150.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.822 Å | ||||||||||||||||||||||||||||||||||||
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Half map: #1
| File | emd_74150_half_map_1.map | ||||||||||||
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| Density Histograms |
-Half map: #2
| File | emd_74150_half_map_2.map | ||||||||||||
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| Density Histograms |
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Sample components
+Entire : Acanthamoeba castellanii OXPHOS complex III intermediate assembly...
+Supramolecule #1: Acanthamoeba castellanii OXPHOS complex III intermediate assembly...
+Macromolecule #1: Peptidase M16 family protein
+Macromolecule #2: Cytochrome b-c1 complex subunit 8
+Macromolecule #3: mitochondrial processing peptidase,Mitochondrial-processing pepti...
+Macromolecule #4: Ubiquinolcytochrome c reductase complex 14 kDa protein
+Macromolecule #5: Cytochrome b-c1 complex subunit 9
+Macromolecule #6: Cytochrome b-c1 complex subunit 6
+Macromolecule #7: Cytochrome c1, heme protein
+Macromolecule #8: Cytochrome b
+Macromolecule #9: Cytochrome b-c1 complex subunit 10
+Macromolecule #10: PROTOPORPHYRIN IX CONTAINING FE
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 2.25 mg/mL | ||||||||||||||
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| Buffer | pH: 7.9 Component:
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| Vitrification | Cryogen name: ETHANE-PROPANE / Chamber humidity: 100 % / Chamber temperature: 277.14 K / Instrument: FEI VITROBOT MARK IV | ||||||||||||||
| Details | Sample was not purified. Instead it was solved from imaging mitochondrial lysate. |
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Electron microscopy
| Microscope | TFS KRIOS |
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| Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
| Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 0.99 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: SPOT SCAN / Imaging mode: DARK FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.6 µm / Nominal magnification: 105000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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About Yorodumi



Keywords
Acanthamoeba castellanii (eukaryote)
Authors
United States, 1 items
Citation















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Processing
FIELD EMISSION GUN
