[English] 日本語
![](img/lk-miru.gif)
- EMDB-5981: Cryo-Electron Microscopy Reconstruction of Glucocorticoid Recepto... -
+
Open data
-
Basic information
Entry | Database: EMDB / ID: EMD-5981 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Cryo-Electron Microscopy Reconstruction of Glucocorticoid Receptor-bound Hsp90-Hsp70-Hop Chaperone Complex | |||||||||
![]() | Reconstruction of client-bound Hsp90-Hsp70-Hop chaperone complex | |||||||||
![]() |
| |||||||||
![]() | Hsp90 / Hsp70 / Hop / Glucocorticoid Receptor | |||||||||
Function / homology | ![]() : / Regulation of NPAS4 gene transcription / cellular heat acclimation / regulation of glucocorticoid biosynthetic process / nuclear glucocorticoid receptor activity / negative regulation of inclusion body assembly / : / positive regulation of nucleotide-binding oligomerization domain containing 2 signaling pathway / C3HC4-type RING finger domain binding / steroid hormone binding ...: / Regulation of NPAS4 gene transcription / cellular heat acclimation / regulation of glucocorticoid biosynthetic process / nuclear glucocorticoid receptor activity / negative regulation of inclusion body assembly / : / positive regulation of nucleotide-binding oligomerization domain containing 2 signaling pathway / C3HC4-type RING finger domain binding / steroid hormone binding / positive regulation of microtubule nucleation / PTK6 Expression / ATP-dependent protein disaggregase activity / dynein axonemal particle / neuroinflammatory response / glucocorticoid metabolic process / microglia differentiation / misfolded protein binding / mammary gland duct morphogenesis / maternal behavior / regulation of mitotic spindle assembly / positive regulation of tumor necrosis factor-mediated signaling pathway / astrocyte differentiation / cellular response to interleukin-7 / cellular response to glucocorticoid stimulus / aggresome / protein folding chaperone complex / RND1 GTPase cycle / motor behavior / positive regulation of tau-protein kinase activity / sperm mitochondrial sheath / dATP binding / Scavenging by Class F Receptors / sulfonylurea receptor binding / CTP binding / positive regulation of protein polymerization / vRNP Assembly / UTP binding / sperm plasma membrane / chaperone-mediated autophagy / telomerase holoenzyme complex assembly / adrenal gland development / protein insertion into mitochondrial outer membrane / cellular response to steroid hormone stimulus / regulation of gluconeogenesis / Respiratory syncytial virus genome replication / Rho GDP-dissociation inhibitor binding / Uptake and function of diphtheria toxin / mitochondrial transport / 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 / PIWI-interacting RNA (piRNA) biogenesis / TPR domain binding / non-chaperonin molecular chaperone ATPase / Assembly and release of respiratory syncytial virus (RSV) virions / dendritic growth cone / mRNA catabolic process / regulation of postsynaptic membrane neurotransmitter receptor levels / Sema3A PAK dependent Axon repulsion / regulation of protein ubiquitination / skeletal muscle contraction / chaperone cofactor-dependent protein refolding / Signaling by ERBB2 / telomere maintenance via telomerase / HSF1-dependent transactivation / positive regulation of cell size / response to unfolded protein / protein unfolding / negative regulation of extrinsic apoptotic signaling pathway in absence of ligand / chaperone-mediated protein complex assembly / Regulation of HSF1-mediated heat shock response / HSF1 activation / regulation of protein-containing complex assembly / Attenuation phase / cellular response to unfolded protein / RHOBTB2 GTPase cycle / estrogen response element binding / FOXO-mediated transcription of oxidative stress, metabolic and neuronal genes / cellular response to dexamethasone stimulus / eNOS activation / axonal growth cone / positive regulation of lamellipodium assembly / DNA polymerase binding / nuclear receptor-mediated steroid hormone signaling pathway / core promoter sequence-specific DNA binding / ATP metabolic process / Tetrahydrobiopterin (BH4) synthesis, recycling, salvage and regulation / inclusion body / positive regulation of defense response to virus by host / Loss of Nlp from mitotic centrosomes / Loss of proteins required for interphase microtubule organization from the centrosome / Recruitment of mitotic centrosome proteins and complexes Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 38.0 Å | |||||||||
![]() | Kirschke E / Goswami D / Southworth DR / Griffin PR / Agard DA | |||||||||
![]() | ![]() Title: Glucocorticoid receptor function regulated by coordinated action of the Hsp90 and Hsp70 chaperone cycles. Authors: Elaine Kirschke / Devrishi Goswami / Daniel Southworth / Patrick R Griffin / David A Agard / ![]() Abstract: The glucocorticoid receptor (GR), like many signaling proteins, depends on the Hsp90 molecular chaperone for in vivo function. Although Hsp90 is required for ligand binding in vivo, purified apo GR ...The glucocorticoid receptor (GR), like many signaling proteins, depends on the Hsp90 molecular chaperone for in vivo function. Although Hsp90 is required for ligand binding in vivo, purified apo GR is capable of binding ligand with no enhancement from Hsp90. We reveal that Hsp70, known to facilitate client delivery to Hsp90, inactivates GR through partial unfolding, whereas Hsp90 reverses this inactivation. Full recovery of ligand binding requires ATP hydrolysis on Hsp90 and the Hop and p23 cochaperones. Surprisingly, Hsp90 ATP hydrolysis appears to regulate client transfer from Hsp70, likely through a coupling of the two chaperone's ATP cycles. Such coupling is embodied in contacts between Hsp90 and Hsp70 in the GR:Hsp70:Hsp90:Hop complex imaged by cryoelectron microscopy. Whereas GR released from Hsp70 is aggregation prone, release from Hsp90 protects GR from aggregation and enhances its ligand affinity. Together, this illustrates how coordinated chaperone interactions can enhance stability, function, and regulation. | |||||||||
History |
|
-
Structure visualization
Movie |
![]() |
---|---|
Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
-
Downloads & links
-EMDB archive
Map data | ![]() | 3.7 MB | ![]() | |
---|---|---|---|---|
Header (meta data) | ![]() ![]() | 13.9 KB 13.9 KB | Display Display | ![]() |
Images | ![]() | 73.2 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Similar structure data |
---|
-
Links
EMDB pages | ![]() ![]() |
---|---|
Related items in Molecule of the Month |
-
Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Reconstruction of client-bound Hsp90-Hsp70-Hop chaperone complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.4 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-
Sample components
-Entire : Human chaperones Hsp90 and Hsp70 bound with the human co-chaperon...
Entire | Name: Human chaperones Hsp90 and Hsp70 bound with the human co-chaperone Hop and the ligand binding domain of the human glucocorticoid receptor N-terminally fused with the maltose binding protein |
---|---|
Components |
|
-Supramolecule #1000: Human chaperones Hsp90 and Hsp70 bound with the human co-chaperon...
Supramolecule | Name: Human chaperones Hsp90 and Hsp70 bound with the human co-chaperone Hop and the ligand binding domain of the human glucocorticoid receptor N-terminally fused with the maltose binding protein type: sample / ID: 1000 Details: Complex was purified using size exclusion chromatography. Oligomeric state: One homodimer of Hsp90 bound to one Hsp70, one Hop, and one MBP-GRLBD Number unique components: 4 |
---|---|
Molecular weight | Theoretical: 380 KDa |
-Macromolecule #1: Heat shock protein 90-alpha
Macromolecule | Name: Heat shock protein 90-alpha / type: protein_or_peptide / ID: 1 / Name.synonym: Hsp90 / Number of copies: 1 / Oligomeric state: Dimer / Recombinant expression: Yes |
---|---|
Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 84.66 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | UniProtKB: Heat shock protein HSP 90-alpha / InterPro: Heat shock protein Hsp90 family |
-Macromolecule #2: Heat shock 70 kDa protein
Macromolecule | Name: Heat shock 70 kDa protein / type: protein_or_peptide / ID: 2 / Name.synonym: Hsp70 / Number of copies: 1 / Oligomeric state: monomer / Recombinant expression: Yes |
---|---|
Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 70.052 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | UniProtKB: Heat shock 70 kDa protein 1B / InterPro: Heat shock protein 70 family |
-Macromolecule #3: Glucocorticoid Receptor Ligand Binding Domain
Macromolecule | Name: Glucocorticoid Receptor Ligand Binding Domain / type: protein_or_peptide / ID: 3 / Name.synonym: GRLBD Details: Maltose Binding Protein (MBP) tag attached to N-terminus Number of copies: 1 / Oligomeric state: monomer / Recombinant expression: Yes |
---|---|
Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 74.926 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | UniProtKB: Glucocorticoid receptor / InterPro: Glucocorticoid receptor |
-Macromolecule #4: Stress-induced-phosphoprotein 1
Macromolecule | Name: Stress-induced-phosphoprotein 1 / type: protein_or_peptide / ID: 4 / Name.synonym: Hop / Number of copies: 1 / Oligomeric state: monomer / Recombinant expression: Yes |
---|---|
Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 62.639 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | UniProtKB: Stress-induced-phosphoprotein 1 / InterPro: Heat shock chaperonin-binding |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
![]() | single particle reconstruction |
Aggregation state | particle |
-
Sample preparation
Buffer | pH: 7.5 Details: 50 mM HEPES, 50 mM KCl, 2 mM DTT, 5 mM MgCl2, 0.2 mM ADP |
---|---|
Grid | Details: C-flat holey carbon grid, 2 micron hole, 0.5 micron space |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 70 % / Chamber temperature: 75 K / Instrument: FEI VITROBOT MARK I / Method: two one-second blots before plunging |
-
Electron microscopy
Microscope | FEI TECNAI F20 |
---|---|
Alignment procedure | Legacy - Astigmatism: Astigmatism was corrected at 250,000 times magnification. |
Date | Sep 21, 2011 |
Image recording | Category: CCD / Film or detector model: TVIPS TEMCAM-F816 (8k x 8k) / Number real images: 214 / Average electron dose: 30 e/Å2 |
Electron beam | Acceleration voltage: 120 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.2 mm / Nominal defocus max: 4.0 µm / Nominal defocus min: 2.2 µm / Nominal magnification: 60000 |
Sample stage | Specimen holder model: OTHER |
Experimental equipment | ![]() Model: Tecnai F20 / Image courtesy: FEI Company |
-
Image processing
Details | The particles were manually selected. |
---|---|
CTF correction | Details: CTF Find |
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 38.0 Å / Resolution method: OTHER / Software - Name: EMAN / Number images used: 10149 |
Final angle assignment | Details: EMAN: 15 |