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Open data
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Basic information
| Entry | Database: PDB / ID: 7u4t | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Title | Human V-ATPase in state 2 with SidK and mEAK-7 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Keywords | MEMBRANE PROTEIN / V-ATPase / mEAK-7 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Function / homology | Function and homology informationnegative regulation of cellular response to oxidative stress / Blockage of phagosome acidification / proton-transporting two-sector ATPase complex / Ion channel transport / plasma membrane proton-transporting V-type ATPase complex / positive regulation of protein localization to lysosome / Regulation of MITF-M-dependent genes involved in lysosome biogenesis and autophagy / intracellular pH reduction / symbiont-mediated suppression of host phagosome acidification / Nef Mediated CD8 Down-regulation ...negative regulation of cellular response to oxidative stress / Blockage of phagosome acidification / proton-transporting two-sector ATPase complex / Ion channel transport / plasma membrane proton-transporting V-type ATPase complex / positive regulation of protein localization to lysosome / Regulation of MITF-M-dependent genes involved in lysosome biogenesis and autophagy / intracellular pH reduction / symbiont-mediated suppression of host phagosome acidification / Nef Mediated CD8 Down-regulation / eye pigmentation / central nervous system maturation / ATPase-coupled ion transmembrane transporter activity / rostrocaudal neural tube patterning / positive regulation of transforming growth factor beta1 production / Golgi lumen acidification / synaptic vesicle lumen acidification / vacuolar transport / proton-transporting V-type ATPase, V0 domain / extrinsic component of synaptic vesicle membrane / Transferrin endocytosis and recycling / cellular response to increased oxygen levels / vacuolar proton-transporting V-type ATPase, V1 domain / endosome to plasma membrane protein transport / vacuolar proton-transporting V-type ATPase, V0 domain / clathrin-coated vesicle membrane / lysosomal lumen acidification / endosomal lumen acidification / XBP1(S) activates chaperone genes / proton-transporting V-type ATPase complex / Amino acids regulate mTORC1 / head morphogenesis / protein localization to cilium / vacuolar proton-transporting V-type ATPase complex / osteoclast development / Nef Mediated CD4 Down-regulation / ROS and RNS production in phagocytes / vacuolar acidification / regulation of cellular pH / dendritic spine membrane / azurophil granule membrane / microvillus / proton transmembrane transporter activity / TOR signaling / ATPase activator activity / regulation of MAPK cascade / autophagosome membrane / tertiary granule membrane / ficolin-1-rich granule membrane / proton-transporting ATPase activity, rotational mechanism / cilium assembly / RHOA GTPase cycle / positive regulation of Wnt signaling pathway / regulation of macroautophagy / response to amino acid / transporter activator activity / H+-transporting two-sector ATPase / ATP metabolic process / specific granule membrane / Metabolism of Angiotensinogen to Angiotensins / angiotensin maturation / Insulin receptor recycling / enzyme regulator activity / ruffle / receptor-mediated endocytosis of virus by host cell / proton-transporting ATP synthase activity, rotational mechanism / axon terminus / endoplasmic reticulum-Golgi intermediate compartment membrane / RNA endonuclease activity / proton transmembrane transport / receptor-mediated endocytosis / regulation of cell migration / secretory granule / secretory granule membrane / response to nutrient levels / response to insulin / small GTPase binding / transmembrane transport / phagocytic vesicle membrane / endocytosis / apical part of cell / synaptic vesicle membrane / melanosome / positive regulation of canonical Wnt signaling pathway / signaling receptor activity / regulation of cell population proliferation / ATPase binding / response to oxidative stress / Hydrolases; Acting on ester bonds / intracellular iron ion homeostasis / postsynaptic membrane / early endosome / lysosome / endosome / endosome membrane / nuclear speck / cilium / apical plasma membrane / Golgi membrane / axon Similarity search - Function | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Biological species | ![]() Homo sapiens (human) | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Authors | Wang, L. / Fu, T.M. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Funding support | 1items
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Citation | Journal: Proc Natl Acad Sci U S A / Year: 2022Title: Identification of mEAK-7 as a human V-ATPase regulator via cryo-EM data mining. Authors: Longfei Wang / Di Wu / Carol V Robinson / Tian-Min Fu / ![]() Abstract: Vacuolar-type adenosine triphosphatases (V-ATPases) not only function as rotary proton pumps in cellular organelles but also serve as signaling hubs. To identify the endogenous binding partners of V- ...Vacuolar-type adenosine triphosphatases (V-ATPases) not only function as rotary proton pumps in cellular organelles but also serve as signaling hubs. To identify the endogenous binding partners of V-ATPase, we collected a large dataset of human V-ATPases and did extensive classification and focused refinement of human V-ATPases. Unexpectedly, about 17% of particles in state 2 of human V-ATPases display additional density with an overall resolution of 3.3 Å. Structural analysis combined with artificial intelligence modeling enables us to identify this additional density as mEAK-7, a protein involved in mechanistic target of rapamycin (mTOR) signaling in mammals. Our structure shows that mEAK-7 interacts with subunits A, B, D, and E of V-ATPases in state 2. Thus, we propose that mEAK-7 may regulate V-ATPase function through binding to V-ATPases in state 2 as well as mediate mTOR signaling. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 7u4t.cif.gz | 1.6 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb7u4t.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 7u4t.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 7u4t_validation.pdf.gz | 1.8 MB | Display | wwPDB validaton report |
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| Full document | 7u4t_full_validation.pdf.gz | 2 MB | Display | |
| Data in XML | 7u4t_validation.xml.gz | 237.5 KB | Display | |
| Data in CIF | 7u4t_validation.cif.gz | 376.2 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/u4/7u4t ftp://data.pdbj.org/pub/pdb/validation_reports/u4/7u4t | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 26334MC M: map data used to model this data C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
| EM raw data | EMPIAR-11132 (Title: Cryo-EM structures of human V-ATPase / Data size: 8.4 TBData #1: Unaligned multi frame micrographs of human V-ATPase in complex with SidK [micrographs - multiframe]) |
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Links
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Assembly
| Deposited unit | ![]()
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Components
-V-type proton ATPase ... , 14 types, 30 molecules ROJIHMLKSU0123456789QCABDEFGNP
| #1: Protein | Mass: 96512.414 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q93050 | ||||||||||||||||||||||
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| #2: Protein | Mass: 43999.500 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P21283 | ||||||||||||||||||||||
| #3: Protein | Mass: 26183.346 Da / Num. of mol.: 3 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P36543#4: Protein | Mass: 13781.547 Da / Num. of mol.: 3 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O75348#5: Protein | | Mass: 9380.329 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O15342#7: Protein | | Mass: 52067.480 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15904#9: Protein | | Mass: 21418.213 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q99437#10: Protein | Mass: 15743.655 Da / Num. of mol.: 9 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P27449#11: Protein | | Mass: 40369.949 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P61421#12: Protein | Mass: 68379.875 Da / Num. of mol.: 3 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human)References: UniProt: P38606, H+-transporting two-sector ATPase #13: Protein | Mass: 56561.500 Da / Num. of mol.: 3 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P21281#15: Protein | | Mass: 28311.918 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9Y5K8#16: Protein | | Mass: 13388.210 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q16864#17: Protein | | Mass: 55949.949 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9UI12 |
-Protein , 4 types, 6 molecules TVZXYW
| #6: Protein | Mass: 15435.220 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human)References: UniProt: Q6P5S7, Hydrolases; Acting on ester bonds | ||
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| #8: Protein | Mass: 39045.855 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O75787 | ||
| #14: Protein | Mass: 65505.297 Da / Num. of mol.: 3 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() #18: Protein | | Mass: 51056.520 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q6P9B6 |
-Sugars / Non-polymers , 2 types, 9 molecules 


| #19: Sugar | ChemComp-NAG / #20: Chemical | ChemComp-ADP / | |
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-Details
| Has ligand of interest | N |
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| Has protein modification | Y |
-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
| Component | Name: V-ATPase with SidK and mEAK-7 / Type: COMPLEX / Entity ID: #1-#4, #6, #8-#18 / Source: NATURAL |
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| Source (natural) | Organism: Homo sapiens (human) |
| Buffer solution | pH: 7.4 |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Vitrification | Cryogen name: ETHANE |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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| Microscopy | Model: FEI TITAN KRIOS |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2500 nm / Nominal defocus min: 1200 nm |
| Image recording | Electron dose: 50 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
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Processing
| Software | Name: PHENIX / Version: 1.19_4092: / Classification: refinement | ||||||||||||||||||||||||
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| EM software | Name: PHENIX / Category: model refinement | ||||||||||||||||||||||||
| CTF correction | Type: NONE | ||||||||||||||||||||||||
| 3D reconstruction | Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 170000 / Symmetry type: POINT | ||||||||||||||||||||||||
| Refine LS restraints |
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FIELD EMISSION GUN