|Entry||Database: PDB / ID: 6qjm|
|Title||Cryo-EM structure of heparin-induced 2N4R tau twister filaments|
|Components||Microtubule-associated protein tauTau protein|
|Keywords||PROTEIN FIBRIL / Recombinant tau protein / heparin / filament / cross-beta structure|
|Function / homology||Tau and MAP proteins tubulin-binding repeat signature. / Activation of AMPK downstream of NMDARs / Caspase-mediated cleavage of cytoskeletal proteins / Tau and MAP proteins tubulin-binding repeat profile. / Tau and MAP protein, tubulin-binding repeat / Microtubule-associated protein Tau / Microtubule associated protein, tubulin-binding repeat / axonal transport / plus-end-directed organelle transport along microtubule / positive regulation of diacylglycerol kinase activity ...Tau and MAP proteins tubulin-binding repeat signature. / Activation of AMPK downstream of NMDARs / Caspase-mediated cleavage of cytoskeletal proteins / Tau and MAP proteins tubulin-binding repeat profile. / Tau and MAP protein, tubulin-binding repeat / Microtubule-associated protein Tau / Microtubule associated protein, tubulin-binding repeat / axonal transport / plus-end-directed organelle transport along microtubule / positive regulation of diacylglycerol kinase activity / neurofibrillary tangle assembly / histone-dependent DNA binding / negative regulation of establishment of protein localization to mitochondrion / neurofibrillary tangle / regulation of chromosome organization / positive regulation of protein localization to synapse / phosphatidylinositol bisphosphate binding / rRNA metabolic process / tubulin complex / microtubule lateral binding / generation of neurons / negative regulation of mitochondrial fission / regulation of long-term synaptic depression / negative regulation of mitochondrial membrane potential / lipoprotein particle binding / microtubule polymerization / axonal transport of mitochondrion / intracellular distribution of mitochondria / axon development / positive regulation of cellular protein localization / negative regulation of tubulin deacetylation / AT DNA binding / regulation of mitochondrial fission / central nervous system neuron development / glial cell projection / regulation of calcium-mediated signaling / negative regulation of kinase activity / internal protein amino acid acetylation / somatodendritic compartment / regulation of microtubule polymerization / dynactin binding / stress granule assembly / receptor ligand activity / cellular response to brain-derived neurotrophic factor stimulus / axolemma / supramolecular fiber organization / cytoplasmic microtubule organization / main axon / regulation of microtubule polymerization or depolymerization / regulation of response to DNA damage stimulus / positive regulation of axon extension / regulation of microtubule cytoskeleton organization / apolipoprotein binding / positive regulation of microtubule polymerization / axon cytoplasm / synapse assembly / phosphatidylinositol binding / amyloid fibril formation / microglial cell activation / nuclear periphery / synapse organization / regulation of autophagy / regulation of synaptic plasticity / astrocyte activation / cellular response to nerve growth factor stimulus / cytoplasmic ribonucleoprotein granule / protein phosphatase 2A binding / positive regulation of superoxide anion generation / Hsp90 protein binding / microtubule cytoskeleton organization / protein binding, bridging / memory / cellular response to reactive oxygen species / microtubule cytoskeleton / cell body / regulation of cellular response to heat / neuron projection development / single-stranded DNA binding / activation of cysteine-type endopeptidase activity involved in apoptotic process / response to lead ion / positive regulation of neuron death / SH3 domain binding / learning or memory / growth cone / intracellular / actin binding / cell-cell signaling / cellular response to heat / double-stranded DNA binding / chaperone binding / protein complex oligomerization / microtubule / microtubule binding / protein homooligomerization / dendritic spine / negative regulation of gene expression / nuclear speck / sequence-specific DNA binding / membrane raft / neuron projection|
Function and homology information
|Specimen source||Homo sapiens (human)|
|Method||ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / 3.3 Å resolution|
|Authors||Zhang, W. / Falcon, B. / Murzin, A.G. / Fan, J. / Crowther, R.A. / Goedert, M. / Scheres, S.H.W.|
|Funding support||United Kingdom , 2 items |
|Citation||Journal: Elife / Year: 2019|
Title: Heparin-induced tau filaments are polymorphic and differ from those in Alzheimer's and Pick's diseases.
Authors: Wenjuan Zhang / Benjamin Falcon / Alexey G Murzin / Juan Fan / R Anthony Crowther / Michel Goedert / Sjors Hw Scheres
Abstract: Assembly of microtubule-associated protein tau into filamentous inclusions underlies a range of neurodegenerative diseases. Tau filaments adopt different conformations in Alzheimer's and Pick's ...Assembly of microtubule-associated protein tau into filamentous inclusions underlies a range of neurodegenerative diseases. Tau filaments adopt different conformations in Alzheimer's and Pick's diseases. Here, we used cryo- and immuno- electron microscopy to characterise filaments that were assembled from recombinant full-length human tau with four (2N4R) or three (2N3R) microtubule-binding repeats in the presence of heparin. 2N4R tau assembles into multiple types of filaments, and the structures of three types reveal similar 'kinked hairpin' folds, in which the second and third repeats pack against each other. 2N3R tau filaments are structurally homogeneous, and adopt a dimeric core, where the third repeats of two tau molecules pack in a parallel manner. The heparin-induced tau filaments differ from those of Alzheimer's or Pick's disease, which have larger cores with different repeat compositions. Our results illustrate the structural versatility of amyloid filaments, and raise questions about the relevance of in vitro assembly.
SummaryFull reportAbout validation report
|Date||Deposition: Jan 24, 2019 / Release: Feb 27, 2019|
|Structure viewer||Molecule: |
Downloads & links
A: Microtubule-associated protein tau
B: Microtubule-associated protein tau
C: Microtubule-associated protein tau
Mass: 5189.081 Da / Num. of mol.: 3 / Source: (gene. exp.) Homo sapiens (human) / Gene: MAPT, MAPTL, MTBT1, TAUTau protein / Plasmid name: pRK172 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: P10636
|Experiment||Method: ELECTRON MICROSCOPY|
|EM experiment||Aggregation state: FILAMENT / Reconstruction method: single particle reconstruction|
|Component||Name: heparin-induced 2N4R tau twister filaments / Type: COMPLEX|
Details: Recombinant tau protein was induced into filaments by incubation with heparin at 37 C for 3 days
Entity ID: 1 / Source: RECOMBINANT
|Molecular weight||Value: 45.85 kDa/nm / Experimental value: NO|
|Source (natural)||Organism: Homo sapiens (human)|
|Source (recombinant)||Organism: Escherichia coli BL21(DE3) (bacteria) / Plasmid: PRK172|
|Buffer solution||Details: 20 mM Tris, pH 7.4, 100mM NaCl / pH: 7.4|
|Specimen||Conc.: 2 mg/ml|
Details: Recombinant tau protein was induced into filaments by incubation with heparin at 37 C for 3 days. The filaments were pronase-treated before making Cryo-grids.
Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
|Specimen support||Grid material: GOLD / Grid mesh size: 300 / Grid type: Quantifoil R1.2/1.3|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 kelvins / Details: Blot force: -12 ; Blot time: 4s|
-Electron microscopy imaging
Model: Tecnai Polara / Image courtesy: FEI Company
|Microscopy||Microscope model: FEI POLARA 300|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM|
|Electron lens||Mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 2800 nm / Nominal defocus min: 1700 nm / Cs: 2 mm / C2 aperture diameter: 5 microns / Alignment procedure: COMA FREE|
|Specimen holder||Cryogen: NITROGEN / Specimen holder model: GATAN LIQUID NITROGEN|
|Image recording||Average exposure time: 1 sec. / Electron dose: 48 e/Å2|
Details: Images were collected in movie-mode at 30 frames per second
Detector mode: INTEGRATING / Film or detector model: FEI FALCON III (4k x 4k) / Number of grids imaged: 1 / Number of real images: 717
|Image scans||Sampling size: 14 microns / Width: 4096 / Height: 4096|
|Image processing||Details: Movie frames were gain-corrected, aligned, dose weighted and then summed into a single micrograph using MOTIONCOR2 (Zheng et al., 2017)|
|CTF correction||Details: Aligned, non-dose-weighted micrographs were used to estimate the contrast transfer function (CTF) using CTFFIND4.1|
Type: PHASE FLIPPING AND AMPLITUDE CORRECTION
|Helical symmerty||Angular rotation/subunit: -3.38 deg. / Axial rise/subunit: 4.7 Å / Axial symmetry: C1|
|Particle selection||Details: Manually picked / Number of particles selected: 187555|
|3D reconstruction||Resolution: 3.3 Å / Resolution method: FSC 0.143 CUT-OFF / Number of particles: 141461 / Algorithm: FOURIER SPACE / Details: we performed two rounds of 3D auto-refinement. / Number of class averages: 6 / Symmetry type: HELICAL|
|Atomic model building||Details: A stack of three consecutive monomers was refined to preserve nearest-neighbour interactions for the middle chain. Side-chain clashes were detected using MOLPROBITY, and corrected by iterative cycles of real-space refinement in COOT and Fourier-space refinement in REFMAC and PHENIX. For each refined structure, separate model refinements were performed against a single half-map, and the resulting model was compared to the other half-map to confirm the absence of overfitting.|
Overall b value: 58.51 / Ref protocol: AB INITIO MODEL / Ref space: RECIPROCAL / Target criteria: Fourier shell correlation
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