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- PDB-3j4r: Pseudo-atomic model of the AKAP18-PKA Complex in a linear conform... -
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Basic information
Entry | Database: PDB / ID: 3j4r | ||||||
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Title | Pseudo-atomic model of the AKAP18-PKA Complex in a linear conformation derived from electron microscopy | ||||||
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![]() | TRANSFERASE / A-kinase anchoring protein / cAMP-Dependent Kinase / RII / PKA regulatory subunit II / phosphorylation / anchoring / intrinsic disorder | ||||||
Function / homology | ![]() spontaneous exocytosis of neurotransmitter / PKA activation in glucagon signalling / CREB1 phosphorylation through the activation of Adenylate Cyclase / negative regulation of meiotic cell cycle / HDL assembly / DARPP-32 events / Rap1 signalling / PKA activation / Vasopressin regulates renal water homeostasis via Aquaporins / Regulation of insulin secretion ...spontaneous exocytosis of neurotransmitter / PKA activation in glucagon signalling / CREB1 phosphorylation through the activation of Adenylate Cyclase / negative regulation of meiotic cell cycle / HDL assembly / DARPP-32 events / Rap1 signalling / PKA activation / Vasopressin regulates renal water homeostasis via Aquaporins / Regulation of insulin secretion / GPER1 signaling / Hedgehog 'off' state / Glucagon-like Peptide-1 (GLP1) regulates insulin secretion / regulation of membrane repolarization / exocytic vesicle / Loss of Nlp from mitotic centrosomes / Recruitment of mitotic centrosome proteins and complexes / Loss of proteins required for interphase microtubule organization from the centrosome / Recruitment of NuMA to mitotic centrosomes / Anchoring of the basal body to the plasma membrane / MAPK6/MAPK4 signaling / GLI3 is processed to GLI3R by the proteasome / AURKA Activation by TPX2 / Factors involved in megakaryocyte development and platelet production / cAMP-dependent protein kinase regulator activity / Regulation of PLK1 Activity at G2/M Transition / Interleukin-3, Interleukin-5 and GM-CSF signaling / CD209 (DC-SIGN) signaling / Mitochondrial protein degradation / positive regulation of potassium ion transmembrane transport / RET signaling / nucleotide-activated protein kinase complex / regulation of protein kinase A signaling / Ion homeostasis / VEGFA-VEGFR2 Pathway / regulation of cellular respiration / regulation of protein processing / protein localization to lipid droplet / regulation of bicellular tight junction assembly / cellular response to parathyroid hormone stimulus / cAMP-dependent protein kinase inhibitor activity / cAMP-dependent protein kinase / cellular response to cold / protein kinase A binding / regulation of osteoblast differentiation / sperm capacitation / cAMP-dependent protein kinase activity / ciliary base / negative regulation of glycolytic process through fructose-6-phosphate / cAMP-dependent protein kinase complex / AMP-activated protein kinase activity / postsynaptic modulation of chemical synaptic transmission / cellular response to glucagon stimulus / AMP binding / protein kinase A regulatory subunit binding / plasma membrane raft / protein kinase A catalytic subunit binding / axoneme / mesoderm formation / lateral plasma membrane / sperm flagellum / small molecule binding / negative regulation of smoothened signaling pathway / regulation of proteasomal protein catabolic process / beta-2 adrenergic receptor binding / cAMP binding / cellular response to cAMP / regulation of synaptic transmission, glutamatergic / positive regulation of gluconeogenesis / sperm midpiece / negative regulation of TORC1 signaling / T-tubule / protein kinase A signaling / protein serine/threonine/tyrosine kinase activity / protein export from nucleus / hippocampal mossy fiber to CA3 synapse / positive regulation of protein export from nucleus / acrosomal vesicle / sarcoplasmic reticulum / neural tube closure / cellular response to glucose stimulus / regulation of protein phosphorylation / modulation of chemical synaptic transmission / neuromuscular junction / adenylate cyclase-activating G protein-coupled receptor signaling pathway / positive regulation of insulin secretion / mRNA processing / small GTPase binding / presynapse / kinase activity / cellular response to heat / manganese ion binding / postsynapse / peptidyl-serine phosphorylation / dendritic spine / regulation of cell cycle / protein kinase activity / nuclear speck / apical plasma membrane / protein domain specific binding Similarity search - Function | ||||||
Biological species | ![]() ![]() ![]() | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / negative staining / Resolution: 35 Å | ||||||
![]() | Reichow, S.L. / Gonen, T. | ||||||
![]() | ![]() Title: Intrinsic disorder within an AKAP-protein kinase A complex guides local substrate phosphorylation. Authors: F Donelson Smith / Steve L Reichow / Jessica L Esseltine / Dan Shi / Lorene K Langeberg / John D Scott / Tamir Gonen / ![]() Abstract: Anchoring proteins sequester kinases with their substrates to locally disseminate intracellular signals and avert indiscriminate transmission of these responses throughout the cell. Mechanistic ...Anchoring proteins sequester kinases with their substrates to locally disseminate intracellular signals and avert indiscriminate transmission of these responses throughout the cell. Mechanistic understanding of this process is hampered by limited structural information on these macromolecular complexes. A-kinase anchoring proteins (AKAPs) spatially constrain phosphorylation by cAMP-dependent protein kinases (PKA). Electron microscopy and three-dimensional reconstructions of type-II PKA-AKAP18γ complexes reveal hetero-pentameric assemblies that adopt a range of flexible tripartite configurations. Intrinsically disordered regions within each PKA regulatory subunit impart the molecular plasticity that affords an ∼16 nanometer radius of motion to the associated catalytic subunits. Manipulating flexibility within the PKA holoenzyme augmented basal and cAMP responsive phosphorylation of AKAP-associated substrates. Cell-based analyses suggest that the catalytic subunit remains within type-II PKA-AKAP18γ complexes upon cAMP elevation. We propose that the dynamic movement of kinase sub-structures, in concert with the static AKAP-regulatory subunit interface, generates a solid-state signaling microenvironment for substrate phosphorylation. DOI: http://dx.doi.org/10.7554/eLife.01319.001. | ||||||
History |
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Structure visualization
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Structure viewer | Molecule: ![]() ![]() |
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PDBx/mmCIF format | ![]() | 323.9 KB | Display | ![]() |
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PDB format | ![]() | 259 KB | Display | ![]() |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
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-Validation report
Summary document | ![]() | 712.6 KB | Display | ![]() |
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Full document | ![]() | 772.4 KB | Display | |
Data in XML | ![]() | 51 KB | Display | |
Data in CIF | ![]() | 76.3 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 5756MC ![]() 5755C ![]() 3j4qC 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: 39478.773 Da / Num. of mol.: 1 / Fragment: SEE REMARK 999 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() | ||||
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#2: Protein | Mass: 45644.266 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() ![]() #3: Protein | Mass: 40628.555 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() ![]() Sequence details | AKAP18 IN THIS ENTRY IS FROM HOMO SAPIENS, BUT THE MODELED SEQUENCE IS FROM RATTUS NORVEGICUS | |
-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
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Buffer solution | Name: 25 mM HEPES, pH 7.4, 200 mM NaCl, 0.5 mM EDTA, 1 mM DTT pH: 7.4 Details: 25 mM HEPES, pH 7.4, 200 mM NaCl, 0.5 mM EDTA, 1 mM DTT | |||||||||||||||||||||||||
Specimen | Conc.: 0.005 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: YES / Vitrification applied: NO / Details: Stain 0.75% (w/v) uranyl formate | |||||||||||||||||||||||||
EM staining | Type: NEGATIVE / Material: Uranyl Formate | |||||||||||||||||||||||||
Specimen support | Details: 200 mesh copper grid with thin carbon support |
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Electron microscopy imaging
Microscopy | Model: FEI TECNAI 12 / Date: May 1, 2012 |
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Electron gun | Electron source: LAB6 / Accelerating voltage: 120 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 52000 X / Nominal defocus max: 2500 nm / Nominal defocus min: 1000 nm / Cs: 6.3 mm Astigmatism: Objective lens astigmatism was corrected at 100,000 times magnification Camera length: 0 mm |
Specimen holder | Specimen holder model: OTHER / Specimen holder type: Single-Tilt / Temperature: 298 K / Tilt angle max: 50 ° / Tilt angle min: 0 ° |
Image recording | Electron dose: 15 e/Å2 / Film or detector model: GENERIC GATAN |
Image scans | Num. digital images: 1335 |
Radiation | Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray |
Radiation wavelength | Relative weight: 1 |
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Processing
EM software |
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CTF correction | Details: Each Micrograph | ||||||||||||||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||||||||||
3D reconstruction | Method: Angular Reconstitution and Refinement / Resolution: 35 Å / Resolution method: FSC 0.5 CUT-OFF / Num. of particles: 1000 / Nominal pixel size: 4.2 Å / Actual pixel size: 4.2 Å Details: (Single particle details: Images were processed in IMAGIC and ISAC. 3D reconstruction was done in IMAGIC and FREALIGN.) (Single particle--Applied symmetry: C1) Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||
Atomic model building |
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Atomic model building | Source name: PDB / Type: experimental model
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Refinement step | Cycle: LAST
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