|Entry||Database: PDB / ID: 6mg8|
|Title||Structural basis for cholesterol transport-like activity of the Hedgehog receptor Patched|
|Components||Protein patched homolog 1|
|Keywords||MEMBRANE PROTEIN / Receptor Membrane protein|
|Function / homology||Transmembrane receptor, patched / Hedgehog 'on' state / Hedgehog 'off' state / Ligand-receptor interactions / Activation of SMO / Sterol-sensing domain / Patched family / Sterol-sensing domain of SREBP cleavage-activation / Protein patched/dispatched / Sterol-sensing domain (SSD) profile. ...Transmembrane receptor, patched / Hedgehog 'on' state / Hedgehog 'off' state / Ligand-receptor interactions / Activation of SMO / Sterol-sensing domain / Patched family / Sterol-sensing domain of SREBP cleavage-activation / Protein patched/dispatched / Sterol-sensing domain (SSD) profile. / neural plate axis specification / hedgehog receptor activity / response to chlorate / cell proliferation involved in metanephros development / smoothened signaling pathway involved in dorsal/ventral neural tube patterning / cell differentiation involved in kidney development / neural tube patterning / smoothened binding / epidermal cell fate specification / hedgehog family protein binding / hindlimb morphogenesis / neural tube formation / spinal cord motor neuron differentiation / mammary gland duct morphogenesis / dorsal/ventral neural tube patterning / patched binding / limb morphogenesis / somite development / cellular response to cholesterol / negative regulation of smoothened signaling pathway / negative regulation of cell division / keratinocyte proliferation / negative regulation of multicellular organism growth / positive regulation of cholesterol efflux / pharyngeal system development / dorsal/ventral pattern formation / positive regulation of epidermal cell differentiation / mammary gland epithelial cell differentiation / cell fate determination / commissural neuron axon guidance / smoothened signaling pathway / mammary gland development / prostate gland development / regulation of smoothened signaling pathway / embryonic organ development / pattern specification process / embryonic limb morphogenesis / cholesterol binding / negative regulation of osteoblast differentiation / regulation of mitotic cell cycle / regulation of growth / branching involved in ureteric bud morphogenesis / epidermis development / negative regulation of epithelial cell proliferation / dendritic growth cone / heart morphogenesis / axonal growth cone / protein processing / cilium / cyclin binding / protein localization to plasma membrane / response to retinoic acid / regulation of protein localization / liver regeneration / neural tube closure / response to mechanical stimulus / animal organ morphogenesis / caveola / brain development / midbody / negative regulation of DNA-binding transcription factor activity / glucose homeostasis / in utero embryonic development / regulation of cell population proliferation / response to estradiol / heparin binding / postsynaptic density / protein-containing complex binding / response to drug / intracellular membrane-bounded organelle / negative regulation of cell population proliferation / negative regulation of transcription, DNA-templated / positive regulation of transcription, DNA-templated / integral component of plasma membrane / Golgi apparatus / negative regulation of transcription by RNA polymerase II / signal transduction / perinuclear region of cytoplasm / zinc ion binding / extracellular region / plasma membrane / nucleus / Protein patched homolog 1|
Function and homology information
|Specimen source||Mus musculus (house mouse)|
|Method||ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / 3.6 Å resolution|
|Authors||Zhang, Y. / Bulkley, D. / Xin, Y. / Roberts, K.J. / Asarnow, D.E. / Sharma, A. / Myers, B.R. / Cho, W. / Cheng, Y. / Beachy, P.A.|
|Citation||Journal: Cell / Year: 2018|
Title: Structural Basis for Cholesterol Transport-like Activity of the Hedgehog Receptor Patched.
Authors: Yunxiao Zhang / David P Bulkley / Yao Xin / Kelsey J Roberts / Daniel E Asarnow / Ashutosh Sharma / Benjamin R Myers / Wonhwa Cho / Yifan Cheng / Philip A Beachy
Abstract: Hedgehog protein signals mediate tissue patterning and maintenance by binding to and inactivating their common receptor Patched, a 12-transmembrane protein that otherwise would suppress the activity ...Hedgehog protein signals mediate tissue patterning and maintenance by binding to and inactivating their common receptor Patched, a 12-transmembrane protein that otherwise would suppress the activity of the 7-transmembrane protein Smoothened. Loss of Patched function, the most common cause of basal cell carcinoma, permits unregulated activation of Smoothened and of the Hedgehog pathway. A cryo-EM structure of the Patched protein reveals striking transmembrane domain similarities to prokaryotic RND transporters. A central hydrophobic conduit with cholesterol-like contents courses through the extracellular domain and resembles that used by other RND proteins to transport substrates, suggesting Patched activity in cholesterol transport. Cholesterol activity in the inner leaflet of the plasma membrane is reduced by PTCH1 expression but rapidly restored by Hedgehog stimulation, suggesting that PTCH1 regulates Smoothened by controlling cholesterol availability.
SummaryFull reportAbout validation report
|Date||Deposition: Sep 13, 2018 / Release: Nov 28, 2018|
|Structure viewer||Molecule: |
Downloads & links
A: Protein patched homolog 1
|#1: Protein/peptide|| |
Mass: 145357.844 Da / Num. of mol.: 1 / Source: (gene. exp.) Mus musculus (house mouse) / Gene: Ptch1, Ptch / Cell line (production host): HEK293 / Production host: Homo sapiens (human) / References: UniProt: Q61115
Mass: 197.236 Da / Num. of mol.: 4
|Nonpolymer details||THE AUTHORS STATE THAT THE EXACT IDENTITY OF THE LIGAND IS UNKNOWN. CHOLESTEROL WAS MODELED INTO ...THE AUTHORS STATE THAT THE EXACT IDENTITY OF THE LIGAND IS UNKNOWN. CHOLESTERO|
|Experiment||Method: ELECTRON MICROSCOPY|
|EM experiment||Aggregation state: PARTICLE / Reconstruction method: single particle reconstruction|
|Component||Name: Patched1 protein solubilized in amphipol / Type: COMPLEX / Entity ID: 1 / Source: RECOMBINANT|
|Molecular weight||Value: 0.1452 MDa / Experimental value: NO|
|Source (natural)||Organism: Homo sapiens (human)|
|Source (recombinant)||Cell: HEK293 / Organism: Homo sapiens (human) / Plasmid: BacMam|
|Buffer solution||pH: 7.4|
|Specimen||Conc.: 0.3 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES|
|Specimen support||Grid material: COPPER / Grid mesh size: 400 / Grid type: Quantifoil R1.2/1.3|
|Vitrification||Instrument: FEI VITROBOT MARK III / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 23 kelvins|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Microscopy||Microscope model: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM|
|Electron lens||Mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm|
|Image recording||Average exposure time: 8 sec. / Electron dose: 38 e/Å2 / Detector mode: SUPER-RESOLUTION / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Number of grids imaged: 1 / Number of real images: 5236|
|Image scans||Movie frames/image: 40|
|Software||Name: PHENIX / Version: dev_2608: / Classification: refinement|
|CTF correction||Type: PHASE FLIPPING AND AMPLITUDE CORRECTION|
|Particle selection||Details: Number of particles selected after rough initial 2D classification|
Number of particles selected: 378828
|Symmetry||Point symmetry: C1|
|3D reconstruction||Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Number of particles: 245725 / Symmetry type: POINT|
|Atomic model building||Ref protocol: AB INITIO MODEL / Ref space: REAL|
|Refine LS restraints|
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