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- PDB-8iwm: Cryo-EM structure of the PEA-bound mTAAR9 complex -

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Basic information

Entry
Database: PDB / ID: 8iwm
TitleCryo-EM structure of the PEA-bound mTAAR9 complex
ComponentsTrace amine-associated receptor 9
KeywordsMEMBRANE PROTEIN / PEA / mTAAR9
Function / homology
Function and homology information


Amine ligand-binding receptors / trace-amine receptor activity / G alpha (s) signalling events / plasma membrane
Similarity search - Function
Trace amine associated receptor family / Serpentine type 7TM GPCR chemoreceptor Srsx / G-protein coupled receptors family 1 signature. / G protein-coupled receptor, rhodopsin-like / GPCR, rhodopsin-like, 7TM / G-protein coupled receptors family 1 profile. / 7 transmembrane receptor (rhodopsin family)
Similarity search - Domain/homology
2-PHENYLETHYLAMINE / Trace amine-associated receptor 9
Similarity search - Component
Biological speciesMus musculus (house mouse)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.17 Å
AuthorsSun, J.P. / Li, Q. / Yang, F. / Xu, Y.F. / Guo, L.L. / Lian, S. / Zhang, M.H. / Rong, N.K.
Funding support China, 1items
OrganizationGrant numberCountry
National Natural Science Foundation of China (NSFC)2018YFC1003600 China
CitationJournal: Nature / Year: 2023
Title: Structural basis of amine odorant perception by a mammal olfactory receptor.
Authors: Lulu Guo / Jie Cheng / Shuo Lian / Qun Liu / Yan Lu / Yuan Zheng / Kongkai Zhu / Minghui Zhang / Yalei Kong / Chao Zhang / Naikang Rong / Yuming Zhuang / Guoxing Fang / Jingjing Jiang / ...Authors: Lulu Guo / Jie Cheng / Shuo Lian / Qun Liu / Yan Lu / Yuan Zheng / Kongkai Zhu / Minghui Zhang / Yalei Kong / Chao Zhang / Naikang Rong / Yuming Zhuang / Guoxing Fang / Jingjing Jiang / Tianyao Zhang / Xiang Han / Zili Liu / Ming Xia / Shangming Liu / Lei Zhang / Stephen D Liberles / Xiao Yu / Yunfei Xu / Fan Yang / Qian Li / Jin-Peng Sun /
Abstract: Odorants are detected as smell in the nasal epithelium of mammals by two G-protein-coupled receptor families, the odorant receptors and the trace amine-associated receptors (TAARs). TAARs emerged ...Odorants are detected as smell in the nasal epithelium of mammals by two G-protein-coupled receptor families, the odorant receptors and the trace amine-associated receptors (TAARs). TAARs emerged following the divergence of jawed and jawless fish, and comprise a large monophyletic family of receptors that recognize volatile amine odorants to elicit both intraspecific and interspecific innate behaviours such as attraction and aversion. Here we report cryo-electron microscopy structures of mouse TAAR9 (mTAAR9) and mTAAR9-G or mTAAR9-G trimers in complex with β-phenylethylamine, N,N-dimethylcyclohexylamine or spermidine. The mTAAR9 structures contain a deep and tight ligand-binding pocket decorated with a conserved DWY motif, which is essential for amine odorant recognition. In the mTAAR9 structure, a unique disulfide bond connecting the N terminus to ECL2 is required for agonist-induced receptor activation. We identify key structural motifs of TAAR family members for detecting monoamines and polyamines and the shared sequence of different TAAR members that are responsible for recognition of the same odour chemical. We elucidate the molecular basis of mTAAR9 coupling to G and G by structural characterization and mutational analysis. Collectively, our results provide a structural basis for odorant detection, receptor activation and G coupling of an amine olfactory receptor.
History
DepositionMar 30, 2023Deposition site: PDBJ / Processing site: PDBC
Revision 1.0May 31, 2023Provider: repository / Type: Initial release
Revision 1.1Jun 7, 2023Group: Database references / Category: citation / Item: _citation.pdbx_database_id_PubMed / _citation.title
Revision 1.2Jun 14, 2023Group: Database references / Category: citation / citation_author
Item: _citation.journal_volume / _citation.page_first ..._citation.journal_volume / _citation.page_first / _citation.page_last / _citation_author.identifier_ORCID

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
R: Trace amine-associated receptor 9
hetero molecules


Theoretical massNumber of molelcules
Total (without water)39,0502
Polymers38,9281
Non-polymers1221
Water0
1


  • Idetical with deposited unit
  • defined by author
  • Evidence: electron microscopy
TypeNameSymmetry operationNumber
identity operation1_5551

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Components

#1: Protein Trace amine-associated receptor 9 / TaR-9 / Trace amine receptor 9 / mTaar9


Mass: 38928.238 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mus musculus (house mouse) / Gene: Taar9 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: Q5QD04
#2: Chemical ChemComp-PEA / 2-PHENYLETHYLAMINE / Phenethylamine


Mass: 122.188 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C8H12N / Feature type: SUBJECT OF INVESTIGATION / Comment: alkaloid*YM
Has ligand of interestY

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

ComponentName: Cryo-EM structure of the PEA-bound mTAAR9 complex / Type: COMPLEX / Entity ID: #1 / Source: RECOMBINANT
Source (natural)Organism: Mus musculus (house mouse)
Source (recombinant)Organism: Spodoptera frugiperda (fall armyworm)
Buffer solutionpH: 7.5
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationCryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: DIFFRACTION / Nominal defocus max: 2000 nm / Nominal defocus min: 1000 nm
Image recordingElectron dose: 1.875 e/Å2 / Film or detector model: GATAN K3 (6k x 4k)

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Processing

EM softwareName: PHENIX / Version: 1.20.1_4487: / Category: model refinement
CTF correctionType: NONE
3D reconstructionResolution: 3.17 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 463012 / Symmetry type: POINT
Refine LS restraints
Refine-IDTypeDev idealNumber
ELECTRON MICROSCOPYf_bond_d0.0128555
ELECTRON MICROSCOPYf_angle_d0.84111654
ELECTRON MICROSCOPYf_dihedral_angle_d4.4661211
ELECTRON MICROSCOPYf_chiral_restr0.0431354
ELECTRON MICROSCOPYf_plane_restr0.0051480

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