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Open data
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Basic information
Entry | Database: PDB / ID: 7k2v | ||||||
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Title | PIKfyve/Fig4/Vac14 complex centered on PIKfyve - map2 | ||||||
![]() | (1-phosphatidylinositol 3-phosphate 5-kinase) x 2 | ||||||
![]() | LIPID BINDING PROTEIN / Lipid kinase / Lipid phosphatase / protein complex | ||||||
Function / homology | ![]() 1-phosphatidylinositol-3-phosphate 5-kinase / 1-phosphatidylinositol-3-phosphate 5-kinase activity / 1-phosphatidylinositol-5-kinase activity / phosphatidylinositol 5-phosphate metabolic process / 1-phosphatidyl-1D-myo-inositol 3,5-bisphosphate metabolic process / phosphatidylinositol-3,5-bisphosphate 5-phosphatase activity / myelin assembly / Synthesis of PIPs at the late endosome membrane / phagosome-lysosome fusion / 1-phosphatidylinositol-4-phosphate 5-kinase activity ...1-phosphatidylinositol-3-phosphate 5-kinase / 1-phosphatidylinositol-3-phosphate 5-kinase activity / 1-phosphatidylinositol-5-kinase activity / phosphatidylinositol 5-phosphate metabolic process / 1-phosphatidyl-1D-myo-inositol 3,5-bisphosphate metabolic process / phosphatidylinositol-3,5-bisphosphate 5-phosphatase activity / myelin assembly / Synthesis of PIPs at the late endosome membrane / phagosome-lysosome fusion / 1-phosphatidylinositol-4-phosphate 5-kinase activity / Synthesis of PIPs at the early endosome membrane / phagosome maturation / Synthesis of PIPs at the Golgi membrane / peptidyl-serine autophosphorylation / regulation of autophagosome assembly / melanosome organization / phosphatidylinositol biosynthetic process / retrograde transport, endosome to Golgi / regulation of reactive oxygen species biosynthetic process / protein targeting to membrane / vesicle membrane / protein localization to nucleus / receptor-mediated endocytosis of virus by host cell / neutrophil chemotaxis / antigen processing and presentation of exogenous peptide antigen via MHC class II / phagocytic vesicle membrane / late endosome membrane / cell-cell junction / early endosome membrane / cytoplasmic vesicle / eukaryotic translation initiation factor 2alpha kinase activity / 3-phosphoinositide-dependent protein kinase activity / DNA-dependent protein kinase activity / ribosomal protein S6 kinase activity / histone H3S10 kinase activity / histone H2AXS139 kinase activity / histone H3S28 kinase activity / histone H4S1 kinase activity / histone H2BS14 kinase activity / histone H3T3 kinase activity / histone H2AS121 kinase activity / histone H2BS36 kinase activity / histone H3S57 kinase activity / histone H2AT120 kinase activity / AMP-activated protein kinase activity / Rho-dependent protein serine/threonine kinase activity / histone H3T6 kinase activity / histone H2AS1 kinase activity / histone H3T11 kinase activity / histone H3T45 kinase activity / non-specific serine/threonine protein kinase / intracellular signal transduction / endosome membrane / membrane raft / Golgi membrane / protein serine kinase activity / protein serine/threonine kinase activity / perinuclear region of cytoplasm / zinc ion binding / ATP binding / metal ion binding / cytosol Similarity search - Function | ||||||
Biological species | ![]() | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 6.6 Å | ||||||
![]() | Lees, J.A. / Reinisch, K.M. / Li, P. | ||||||
Funding support | ![]()
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![]() | ![]() Title: Insights into Lysosomal PI(3,5)P Homeostasis from a Structural-Biochemical Analysis of the PIKfyve Lipid Kinase Complex. Authors: Joshua A Lees / PeiQi Li / Nikit Kumar / Lois S Weisman / Karin M Reinisch / ![]() Abstract: The phosphoinositide PI(3,5)P, generated exclusively by the PIKfyve lipid kinase complex, is key for lysosomal biology. Here, we explore how PI(3,5)P levels within cells are regulated. We find the ...The phosphoinositide PI(3,5)P, generated exclusively by the PIKfyve lipid kinase complex, is key for lysosomal biology. Here, we explore how PI(3,5)P levels within cells are regulated. We find the PIKfyve complex comprises five copies of the scaffolding protein Vac14 and one copy each of the lipid kinase PIKfyve, generating PI(3,5)P from PI3P and the lipid phosphatase Fig4, reversing the reaction. Fig4 is active as a lipid phosphatase in the ternary complex, whereas PIKfyve within the complex cannot access membrane-incorporated phosphoinositides due to steric constraints. We find further that the phosphoinositide-directed activities of both PIKfyve and Fig4 are regulated by protein-directed activities within the complex. PIKfyve autophosphorylation represses its lipid kinase activity and stimulates Fig4 lipid phosphatase activity. Further, Fig4 is also a protein phosphatase acting on PIKfyve to stimulate its lipid kinase activity, explaining why catalytically active Fig4 is required for maximal PI(3,5)P production by PIKfyve in vivo. | ||||||
History |
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Structure visualization
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Structure viewer | Molecule: ![]() ![]() |
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Downloads & links
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Download
PDBx/mmCIF format | ![]() | 204.8 KB | Display | ![]() |
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PDB format | ![]() | 165.7 KB | Display | ![]() |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
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-Validation report
Summary document | ![]() | 814.1 KB | Display | ![]() |
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Full document | ![]() | 883.8 KB | Display | |
Data in XML | ![]() | 36.2 KB | Display | |
Data in CIF | ![]() | 52.7 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 22647MC ![]() 7k1wC ![]() 7k1yC M: map data used to model this data C: citing same article ( |
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Similar structure data |
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Assembly
Deposited unit | ![]()
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Components
#1: Protein | Mass: 30462.967 Da / Num. of mol.: 1 / Fragment: PIPK domain (UNP residues 1822-2085) Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: Q9Y2I7, 1-phosphatidylinositol-3-phosphate 5-kinase |
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#2: Protein | Mass: 51413.039 Da / Num. of mol.: 1 / Fragment: CCT domain (UNP residues 491-927) Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
-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: PIKfyve/Fig4/Vac14 complex / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT |
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Molecular weight | Value: 4.28 MDa / Experimental value: NO |
Source (natural) | Organism: ![]() |
Source (recombinant) | Organism: ![]() |
Buffer solution | pH: 7.8 |
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: ![]() |
Electron lens | Mode: BRIGHT FIELD / Cs: 2.7 mm |
Image recording | Electron dose: 58.4 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k) |
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Processing
CTF correction | Type: NONE |
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3D reconstruction | Resolution: 6.6 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 19998 / Symmetry type: POINT |