|Entry||Database: PDB / ID: 3j5q|
|Title||Structure of TRPV1 ion channel in complex with DkTx and RTX determined by single particle electron cryo-microscopy|
|Keywords||TRANSPORT PROTEIN/TOXIN / TRPV1 channel / DkTx / RTX / TRANSPORT PROTEIN-TOXIN complex|
|Function / homology||Ankyrin repeat / Ankyrin repeat region circular profile. / Ion transport domain / Transient receptor potential channel / Ankyrin repeat / Transient receptor potential cation channel subfamily V / Transient receptor potential cation channel subfamily V member 1 / Ankyrin repeat-containing domain / Ankyrin repeat-containing domain superfamily / Ion transport protein ...Ankyrin repeat / Ankyrin repeat region circular profile. / Ion transport domain / Transient receptor potential channel / Ankyrin repeat / Transient receptor potential cation channel subfamily V / Transient receptor potential cation channel subfamily V member 1 / Ankyrin repeat-containing domain / Ankyrin repeat-containing domain superfamily / Ion transport protein / Ankyrin repeats (3 copies) / Ankyrin repeat profile. / Transient receptor potential channel, vanilloid 1-4 / TRP channels / response to capsazepine / peptide secretion / detection of temperature stimulus involved in thermoception / temperature-gated ion channel activity / sensory perception of mechanical stimulus / positive regulation of gastric acid secretion / detection of chemical stimulus involved in sensory perception of pain / thermoception / dendritic spine membrane / negative regulation of establishment of blood-brain barrier / smooth muscle contraction involved in micturition / excitatory extracellular ligand-gated ion channel activity / urinary bladder smooth muscle contraction / cellular response to temperature stimulus / diet induced thermogenesis / cellular response to acidic pH / calcium ion import across plasma membrane / fever generation / chloride channel regulator activity / temperature homeostasis / cation transmembrane transporter activity / behavioral response to pain / glutamate secretion / detection of temperature stimulus involved in sensory perception of pain / cation channel activity / calcium-release channel activity / intrinsic component of plasma membrane / response to pain / cellular response to ATP / cellular response to alkaloid / ligand-gated ion channel activity / extracellular ligand-gated ion channel activity / phosphatidylinositol binding / microglial cell activation / calcium channel activity / calcium ion transmembrane transport / cellular response to cytokine stimulus / sensory perception of pain / cellular response to nerve growth factor stimulus / response to organonitrogen compound / response to peptide hormone / calcium ion transport / lipid metabolic process / ion transmembrane transport / cellular response to growth factor stimulus / phosphoprotein binding / cellular response to heat / response to pH / transmembrane signaling receptor activity / cellular response to tumor necrosis factor / positive regulation of cytosolic calcium ion concentration / postsynaptic membrane / protein homotetramerization / response to heat / positive regulation of nitric oxide biosynthetic process / ion channel activity / calmodulin binding / neuron projection / synapse / cell junction / dendrite / neuronal cell body / external side of plasma membrane / inflammatory response / positive regulation of apoptotic process / integral component of plasma membrane / negative regulation of transcription by RNA polymerase II / membrane / integral component of membrane / ATP binding / identical protein binding / plasma membrane / metal ion binding / cytosol / Transient receptor potential cation channel subfamily V member 1|
Function and homology information
|Specimen source||Rattus norvegicus (Norway rat)|
Chilobrachys jingzhao (Chilobrachys guangxiensis)
|Method||ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / 3.8 Å resolution|
|Authors||Liao, M. / Cao, E. / Julius, D. / Cheng, Y.|
|Citation||Journal: Nature / Year: 2013|
Title: TRPV1 structures in distinct conformations reveal activation mechanisms.
Authors: Erhu Cao / Maofu Liao / Yifan Cheng / David Julius
Abstract: Transient receptor potential (TRP) channels are polymodal signal detectors that respond to a wide range of physical and chemical stimuli. Elucidating how these channels integrate and convert ...Transient receptor potential (TRP) channels are polymodal signal detectors that respond to a wide range of physical and chemical stimuli. Elucidating how these channels integrate and convert physiological signals into channel opening is essential to understanding how they regulate cell excitability under normal and pathophysiological conditions. Here we exploit pharmacological probes (a peptide toxin and small vanilloid agonists) to determine structures of two activated states of the capsaicin receptor, TRPV1. A domain (consisting of transmembrane segments 1-4) that moves during activation of voltage-gated channels remains stationary in TRPV1, highlighting differences in gating mechanisms for these structurally related channel superfamilies. TRPV1 opening is associated with major structural rearrangements in the outer pore, including the pore helix and selectivity filter, as well as pronounced dilation of a hydrophobic constriction at the lower gate, suggesting a dual gating mechanism. Allosteric coupling between upper and lower gates may account for rich physiological modulation exhibited by TRPV1 and other TRP channels.
SummaryFull reportAbout validation report
|Date||Deposition: Oct 28, 2013 / Release: Dec 4, 2013|
|Structure viewer||Molecule: |
Downloads & links
D: Transient receptor potential cation channel subfamily V member 1
B: Transient receptor potential cation channel subfamily V member 1
E: Transient receptor potential cation channel subfamily V member 1
G: Transient receptor potential cation channel subfamily V member 1
A: Double-knot toxin
C: Double-knot toxin
F: Double-knot toxin
H: Double-knot toxin
Mass: 68242.156 Da / Num. of mol.: 4 / Fragment: SEE REMARK 999 / Source: (gene. exp.) Rattus norvegicus (Norway rat) / Gene: Trpv1, Vr1, Vr1l / Plasmid name: pFastBac1 / Cell line (production host): HEK293S GnTI / Production host: Homo sapiens (human) / References: UniProt:O35433
Mass: 2656.265 Da / Num. of mol.: 4 / Fragment: SEE REMARK 999
Source: (natural) Chilobrachys jingzhao (Chilobrachys guangxiensis)
|Sequence details||THE TRPV1 CONSTRUCT COMPRISES RESIDUES 110-603 AND 627-764, WITH RESIDUES 604-626 DELETED. RESIDUES ...THE TRPV1 CONSTRUCT COMPRISES RESIDUES 110-603 AND 627-764, WITH RESIDUES 604-626 DELETED. RESIDUES 719-764 ARE NOT MODELED, WITH THE EXCEPTION OF 11 RESIDUES (NUMBERED 752-762 IN THE COORDINATE|
|Experiment||Method: ELECTRON MICROSCOPY|
|EM experiment||Aggregation state: PARTICLE / Reconstruction method: single particle reconstruction|
|Component||Name: Rat TRPV1 in complex with DkTx and resiniferatoxin / Type: COMPLEX / Details: Tetramer|
|Molecular weight||Value: 0.3 deg. / Units: MEGADALTONS / Experimental value: NO|
|Buffer solution||Name: 150 mM NaCl, 20 mM HEPES, 2 mM TCEP / Details: 150 mM NaCl, 20 mM HEPES, 2 mM TCEP / pH: 7.4|
|Specimen||Conc.: 0.3 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES|
|Specimen support||Details: 400 mesh Quantifoil grid|
|Vitrification||Instrument: FEI VITROBOT MARK III / Cryogen name: ETHANE / Temp: 120 K / Humidity: 90 %|
Details: Blot for 6 seconds before plunging into liquid ethane (FEI VITROBOT MARK III)
Method: Blot for 6 sec
-Electron microscopy imaging
Model: Tecnai Polara / Image courtesy: FEI Company
|Microscopy||Microscope model: FEI POLARA 300 / Date: Jan 1, 2013|
Details: Gatan K2 Summit operated in super-resolution counting mode, image recorded with dose fractionation method.
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM|
|Electron lens||Mode: BRIGHT FIELDBright-field microscopy / Nominal magnification: 31000 / Calibrated magnification: 31000 / Nominal defocus max: 3000 nm / Nominal defocus min: 1500 nm / Cs: 2 mm / Camera length: 0 mm|
|Specimen holder||Specimen holder model: OTHER / Specimen holder type: FEI Polara cartridge|
|Image recording||Electron dose: 21 e/Å2|
Details: Operated in super-resolution counting mode, dose fractionation
Film or detector model: GATAN K2 SUMMIT (4k x 4k)
|Image scans||Number digital images: 900|
|Radiation||Diffraction protocol: SINGLE WAVELENGTH / Monochromatic or laue m l: M / Scattering type: x-ray|
|Radiation wavelength||Relative weight: 1|
|EM software||Name: Relion / Category: 3D reconstruction|
|CTF correction||Details: Each particle|
|Symmetry||Point symmetry: C4|
|3D reconstruction||Method: Maximum likelihood / Resolution: 3.8 Å / Resolution method: FSC 0.143 CUT-OFF / Number of particles: 36158 / Nominal pixel size: 1.2156 / Actual pixel size: 1.2156|
Details: The entire ankyrin structure was fitted as a single rigid body. As a result, the first two ankyrin repeats (UNP residues 111-199) are included but are not well defined by the experimental density. Interatomic clashes between the ankyrin repeats and the C-terminal beta sheet are the result of poor geometry due to relatively poor experimental density in these regions. Resolution and averaging of DkTx density was insufficient to provide side chain geometry. The DkTx model was generated as a poly-ALA model based on hanatoxin (PDB entry 1D1H) and docked into the DkTx density with slight modification. (Single particle details: 3D classification, refinement, and reconstruction were performed using RELION) (Single particle--Applied symmetry: C4)
Refinement type: HALF-MAPS REFINED INDEPENDENTLY / Symmetry type: POINT
|Atomic model building||Details: REFINEMENT PROTOCOL--de novo model building DETAILS--The entire ankyrin structure was fitted as a single rigid body. As a result, the first two ankyrin repeats (UNP residues 111-199) are included but are not well defined by the experimental density.|
Ref protocol: AB INITIO MODEL / Ref space: REAL
|Number of atoms included #LAST||Protein: 18264 / Nucleic acid: 0 / Ligand: 0 / Solvent: 0 / Total: 18264|
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