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Open data
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Basic information
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| Title | Sub-tomogram average of the V-ATPase from isolated lysosomes | |||||||||
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Keywords | Proton pump / ATP hydrolysis / native lysosomal membrane / V-ATPase / MEMBRANE PROTEIN | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | subtomogram averaging / cryo EM / Resolution: 24.3 Å | |||||||||
Authors | De Jesus-Perez JJ / Mcveigh B / Mihelc EM / Moiseenkova-Bell VY | |||||||||
| Funding support | United States, 1 items
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Citation | Journal: Nat Commun / Year: 2025Title: Visualization of lysosomal membrane proteins by cryo electron tomography. Authors: Bridget M McVeigh / José J De Jesús-Pérez / Dirk H Siepe / Prerana Gogoi / Shrawan Kumar Mageswaran / Marian Kalocsay / Elaine M Mihelc / Vera Y Moiseenkova-Bell / ![]() Abstract: Lysosomes are essential organelles for cellular homeostasis and signaling, with dysfunction linked to neurological disorders, lysosomal storage diseases, and cancer. While proteomics has advanced our ...Lysosomes are essential organelles for cellular homeostasis and signaling, with dysfunction linked to neurological disorders, lysosomal storage diseases, and cancer. While proteomics has advanced our understanding of lysosomal composition, the structural characterization of lysosomal membrane proteins in their native environment remains a significant challenge. Here, we developed a cryo electron tomography workflow to visualize lysosomal membrane proteins within intact, native lysosomal membranes. We isolated endolysosomes by independently targeting two lysosomal membrane proteins, transient receptor potential mucolipin 1 and transmembrane protein 192, enriching organelles that exhibited the expected morphology and proteomic composition of the endolysosomal system. Sub-tomogram averaging enabled the structural refinement of key membrane and membrane-associated proteins, including V-ATPase, Flotillin, and Clathrin, directly within the lysosomal membrane, revealing their heterogeneous distribution across endolysosomal organelles. By integrating proteomics with structural biology, our workflow establishes a powerful platform for studying lysosomal membrane protein function in health and disease, paving the way for future discoveries in membrane-associated lysosomal mechanisms. | |||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_47080.map.gz | 9.6 MB | EMDB map data format | |
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| Header (meta data) | emd-47080-v30.xml emd-47080.xml | 15.9 KB 15.9 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_47080_fsc.xml | 12.9 KB | Display | FSC data file |
| Images | emd_47080.png | 53.6 KB | ||
| Filedesc metadata | emd-47080.cif.gz | 4.3 KB | ||
| Others | emd_47080_half_map_1.map.gz emd_47080_half_map_2.map.gz | 761.6 MB 761.5 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-47080 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-47080 | HTTPS FTP |
-Validation report
| Summary document | emd_47080_validation.pdf.gz | 913.7 KB | Display | EMDB validaton report |
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| Full document | emd_47080_full_validation.pdf.gz | 913.3 KB | Display | |
| Data in XML | emd_47080_validation.xml.gz | 25 KB | Display | |
| Data in CIF | emd_47080_validation.cif.gz | 31.7 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-47080 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-47080 | HTTPS FTP |
-Related structure data
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Map
| File | Download / File: emd_47080.map.gz / Format: CCP4 / Size: 178 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: 1.69 Å | ||||||||||||||||||||||||||||||||||||
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Half map: #1
| File | emd_47080_half_map_1.map | ||||||||||||
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-Half map: #2
| File | emd_47080_half_map_2.map | ||||||||||||
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Sample components
-Entire : Structure of the V-ATPase from isolated lysosomes
| Entire | Name: Structure of the V-ATPase from isolated lysosomes |
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| Components |
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-Supramolecule #1: Structure of the V-ATPase from isolated lysosomes
| Supramolecule | Name: Structure of the V-ATPase from isolated lysosomes / type: complex / ID: 1 / Parent: 0 |
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| Source (natural) | Organism: Homo sapiens (human) |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | subtomogram averaging |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7.25 |
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| Vitrification | Cryogen name: ETHANE |
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Electron microscopy
| Microscope | TFS KRIOS |
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| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 2.5 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 5.0 µm / Nominal defocus min: 2.0 µm / Nominal magnification: 53000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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About Yorodumi




Keywords
Homo sapiens (human)
Authors
United States, 1 items
Citation

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Processing
FIELD EMISSION GUN

