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Open data
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Basic information
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Title | Hsp90 provides platform for CRaf dephosphorylation by PP5 | |||||||||||||||
![]() | Composite map, used to dock and improve model alignment. | |||||||||||||||
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Function / homology | ![]() regulation of type II interferon-mediated signaling pathway / : / response to arachidonic acid / HSP90-CDC37 chaperone complex / positive regulation of cyclin-dependent protein kinase activity / death-inducing signaling complex assembly / positive regulation of mitophagy in response to mitochondrial depolarization / negative regulation of proteasomal protein catabolic process / Aryl hydrocarbon receptor signalling / peptidyl-serine dephosphorylation ...regulation of type II interferon-mediated signaling pathway / : / response to arachidonic acid / HSP90-CDC37 chaperone complex / positive regulation of cyclin-dependent protein kinase activity / death-inducing signaling complex assembly / positive regulation of mitophagy in response to mitochondrial depolarization / negative regulation of proteasomal protein catabolic process / Aryl hydrocarbon receptor signalling / peptidyl-serine dephosphorylation / peptidyl-threonine dephosphorylation / aryl hydrocarbon receptor complex / dynein axonemal particle / intermediate filament cytoskeleton organization / histone methyltransferase binding / type B pancreatic cell proliferation / regulation of Rho protein signal transduction / SHOC2 M1731 mutant abolishes MRAS complex function / Gain-of-function MRAS complexes activate RAF signaling / positive regulation of protein localization to cell surface / Rap1 signalling / ATP-dependent protein binding / protein kinase regulator activity / regulation of cell motility / response to morphine / protein folding chaperone complex / insulin secretion involved in cellular response to glucose stimulus / negative regulation of protein metabolic process / positive regulation of tau-protein kinase activity / Negative feedback regulation of MAPK pathway / post-transcriptional regulation of gene expression / myosin phosphatase activity / telomerase holoenzyme complex assembly / Respiratory syncytial virus genome replication / regulation of cyclin-dependent protein serine/threonine kinase activity / Uptake and function of diphtheria toxin / GP1b-IX-V activation signalling / IFNG signaling activates MAPKs / protein serine/threonine phosphatase activity / Drug-mediated inhibition of ERBB2 signaling / Resistance of ERBB2 KD mutants to trastuzumab / Resistance of ERBB2 KD mutants to sapitinib / Resistance of ERBB2 KD mutants to tesevatinib / Resistance of ERBB2 KD mutants to neratinib / Resistance of ERBB2 KD mutants to osimertinib / Resistance of ERBB2 KD mutants to afatinib / Resistance of ERBB2 KD mutants to AEE788 / Resistance of ERBB2 KD mutants to lapatinib / Drug resistance in ERBB2 TMD/JMD mutants / protein-serine/threonine phosphatase / TPR domain binding / ERBB2-ERBB3 signaling pathway / positive regulation of transforming growth factor beta receptor signaling pathway / regulation of cell differentiation / Assembly and release of respiratory syncytial virus (RSV) virions / face development / pseudopodium / phosphatase activity / dendritic growth cone / somatic stem cell population maintenance / regulation of type I interferon-mediated signaling pathway / positive regulation of phosphoprotein phosphatase activity / phosphoprotein phosphatase activity / thyroid gland development / neurotrophin TRK receptor signaling pathway / Sema3A PAK dependent Axon repulsion / The NLRP3 inflammasome / regulation of protein ubiquitination / HSF1-dependent transactivation / response to unfolded protein / negative regulation of proteasomal ubiquitin-dependent protein catabolic process / telomere maintenance via telomerase / extrinsic apoptotic signaling pathway via death domain receptors / chaperone-mediated protein complex assembly / MAP kinase kinase kinase activity / HSF1 activation / Attenuation phase / protein targeting / cellular response to interleukin-4 / negative regulation of protein-containing complex assembly / RHOBTB2 GTPase cycle / Purinergic signaling in leishmaniasis infection / Schwann cell development / axonal growth cone / type II interferon-mediated signaling pathway / DNA polymerase binding / supramolecular fiber organization / negative regulation of extrinsic apoptotic signaling pathway via death domain receptors / Signaling by ERBB2 / : / heat shock protein binding / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / activation of adenylate cyclase activity / response to muscle stretch / nitric-oxide synthase regulator activity / myelination / CD209 (DC-SIGN) signaling / Constitutive Signaling by Overexpressed ERBB2 / protein dephosphorylation / ESR-mediated signaling Similarity search - Function | |||||||||||||||
Biological species | ![]() | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.3 Å | |||||||||||||||
![]() | Jaime-Garza M / Nowotny CA / Coutandin D / Wang F / Tabios M / Agard DA | |||||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Hsp90 provides a platform for kinase dephosphorylation by PP5. Authors: Maru Jaime-Garza / Carlos A Nowotny / Daniel Coutandin / Feng Wang / Mariano Tabios / David A Agard / ![]() Abstract: The Hsp90 molecular chaperone collaborates with the phosphorylated Cdc37 cochaperone for the folding and activation of its many client kinases. As with many kinases, the Hsp90 client kinase CRaf is ...The Hsp90 molecular chaperone collaborates with the phosphorylated Cdc37 cochaperone for the folding and activation of its many client kinases. As with many kinases, the Hsp90 client kinase CRaf is activated by phosphorylation at specific regulatory sites. The cochaperone phosphatase PP5 dephosphorylates CRaf and Cdc37 in an Hsp90-dependent manner. Although dephosphorylating Cdc37 has been proposed as a mechanism for releasing Hsp90-bound kinases, here we show that Hsp90 bound kinases sterically inhibit Cdc37 dephosphorylation indicating kinase release must occur before Cdc37 dephosphorylation. Our cryo-EM structure of PP5 in complex with Hsp90:Cdc37:CRaf reveals how Hsp90 both activates PP5 and scaffolds its association with the bound CRaf to dephosphorylate phosphorylation sites neighboring the kinase domain. Thus, we directly show how Hsp90's role in maintaining protein homeostasis goes beyond folding and activation to include post translationally modifying its client kinases. | |||||||||||||||
History |
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Structure visualization
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Downloads & links
-EMDB archive
Map data | ![]() | 59.4 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 31.6 KB 31.6 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() ![]() ![]() ![]() | 11.4 KB 11.4 KB 11.4 KB 11.4 KB | Display Display Display Display | ![]() |
Images | ![]() | 80.9 KB | ||
Others | ![]() ![]() ![]() | 7.2 MB 109.9 MB 103.2 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 740.1 KB | Display | ![]() |
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Full document | ![]() | 739.7 KB | Display | |
Data in XML | ![]() | 18.9 KB | Display | |
Data in CIF | ![]() | 24.1 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8gaeMC ![]() 8gftC C: citing same article ( M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Composite map, used to dock and improve model alignment. | ||||||||||||||||||||
Voxel size | X=Y=Z: 0.835 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: Sharpened composite map, used for detailed model fitting.
File | emd_29895_additional_1.map | ||||||||||||
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Annotation | Sharpened composite map, used for detailed model fitting. | ||||||||||||
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Density Histograms |
-Half map: Composite half map. Used to obtain composite resolution.
File | emd_29895_half_map_1.map | ||||||||||||
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Annotation | Composite half map. Used to obtain composite resolution. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Composite half map. Used to obtain composite resolution.
File | emd_29895_half_map_2.map | ||||||||||||
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Annotation | Composite half map. Used to obtain composite resolution. | ||||||||||||
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Density Histograms |
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Sample components
+Entire : Hsp90:Cdc37:CRaf:PP5 complex
+Supramolecule #1: Hsp90:Cdc37:CRaf:PP5 complex
+Supramolecule #2: Protein Phosphatase 5 (H304A)
+Macromolecule #1: Heat shock protein HSP 90-beta
+Macromolecule #2: Hsp90 co-chaperone Cdc37
+Macromolecule #3: RAF proto-oncogene serine/threonine-protein kinase
+Macromolecule #4: Serine/threonine-protein phosphatase 5
+Macromolecule #5: POTASSIUM ION
+Macromolecule #6: MAGNESIUM ION
+Macromolecule #7: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #8: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #9: MANGANESE (II) ION
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.5 Component:
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Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: GRAPHENE OXIDE / Support film - topology: CONTINUOUS / Support film - Film thickness: 1.0 nm | ||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 283.15 K / Instrument: FEI VITROBOT MARK IV Details: 3UL OF SAMPLE 10C 100% HUMIDITY 30S WAIT TIME 3S BLOT TIME -2 BLOT FORCE. |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Software | Name: SerialEM (ver. 3.8-beta) |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number real images: 4160 / Average electron dose: 69.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.8 µm / Nominal defocus min: 0.8 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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Image processing
-Atomic model buiding 1
Initial model | (Chain: PDB, experimental model, PDB, experimental model, PDB, experimental model) |
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Details | This model was built using rigid body docking in Chimera and ChimeraX for all main chains, and RosettaCM to add remaining fragments not included in previous models. Refinement was done using iterative Phenix and RosettaRelax, and finalized with ISOLDE. |
Refinement | Protocol: RIGID BODY FIT |
Output model | ![]() PDB-8gae: |