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- EMDB-44259: Cryo-EM structure of the mouse TRPM8 channel in complex with the ... -

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Basic information

Entry
Database: EMDB / ID: EMD-44259
TitleCryo-EM structure of the mouse TRPM8 channel in complex with the antagonist TC-I 2014 and the cooling agonist C3
Map dataThe final full map sharpened with a B factor of -30
Sample
  • Complex: Transient receptor potential cation channel subfamily M member 8
    • Protein or peptide: Transient receptor potential cation channel subfamily M member 8
  • Ligand: 3-{7-(trifluoromethyl)-5-[2-(trifluoromethyl)phenyl]-1H-benzimidazol-2-yl}-1-oxa-2-azaspiro[4.5]dec-2-ene
  • Ligand: CHOLESTEROL HEMISUCCINATE
  • Ligand: nonyl(oxo)di(propan-2-yl)-lambda~5~-phosphane
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose
KeywordsTRPM8 / menthol receptor / cold receptor / PI(4 / 5)P2 / cooling agonists / temperature sensing / ion channel / sensory transduction / transient receptor potential ion channel / TRPM8 activation / TRPM8 inhibition / TRPM8 desensitization / TRPM8 antagonists / MEMBRANE PROTEIN
Function / homology
Function and homology information


ligand-gated calcium channel activity / TRP channels / thermoception / response to temperature stimulus / monoatomic ion channel activity / response to cold / calcium ion transmembrane transport / calcium channel activity / intracellular calcium ion homeostasis / calcium ion transport ...ligand-gated calcium channel activity / TRP channels / thermoception / response to temperature stimulus / monoatomic ion channel activity / response to cold / calcium ion transmembrane transport / calcium channel activity / intracellular calcium ion homeostasis / calcium ion transport / positive regulation of cold-induced thermogenesis / membrane raft / external side of plasma membrane / identical protein binding / membrane / plasma membrane
Similarity search - Function
TRPM, SLOG domain / : / SLOG in TRPM / Ion transport domain / Ion transport protein
Similarity search - Domain/homology
Transient receptor potential cation channel subfamily M member 8
Similarity search - Component
Biological speciesMus musculus (house mouse)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.76 Å
AuthorsYin Y / Park C-G / Zhang F / Fedor J / Feng S / Suo Y / Im W / Lee S-Y
Funding support United States, 2 items
OrganizationGrant numberCountry
National Institutes of Health/National Eye Institute (NIH/NEI) United States
National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS) United States
CitationJournal: Sci Adv / Year: 2024
Title: Mechanisms of sensory adaptation and inhibition of the cold and menthol receptor TRPM8.
Authors: Ying Yin / Cheon-Gyu Park / Feng Zhang / Justin G Fedor / Shasha Feng / Yang Suo / Wonpil Im / Seok-Yong Lee /
Abstract: Our sensory adaptation to cold and chemically induced coolness is mediated by the intrinsic property of TRPM8 channels to desensitize. TRPM8 is also implicated in cold-evoked pain disorders and ...Our sensory adaptation to cold and chemically induced coolness is mediated by the intrinsic property of TRPM8 channels to desensitize. TRPM8 is also implicated in cold-evoked pain disorders and migraine, highlighting its inhibitors as an avenue for pain relief. Despite the importance, the mechanisms of TRPM8 desensitization and inhibition remained unclear. We found, using cryo-electron microscopy, electrophysiology, and molecular dynamics simulations, that TRPM8 inhibitors bind selectively to the desensitized state of the channel. These inhibitors were used to reveal the overlapping mechanisms of desensitization and inhibition and that cold and cooling agonists share a common desensitization pathway. Furthermore, we identified the structural determinants crucial for the conformational change in TRPM8 desensitization. Our study illustrates how receptor-level conformational changes alter cold sensation, providing insights into therapeutic development.
History
DepositionMar 25, 2024-
Header (metadata) releaseAug 21, 2024-
Map releaseAug 21, 2024-
UpdateAug 21, 2024-
Current statusAug 21, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_44259.png

Downloads & links

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Map

FileDownload / File: emd_44259.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationThe final full map sharpened with a B factor of -30
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.08 Å/pix.
x 320 pix.
= 345.6 Å		1.08 Å/pix.
x 320 pix.
= 345.6 Å		1.08 Å/pix.
x 320 pix.
= 345.6 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.08 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.1
Minimum - Maximum-1.0622118 - 1.8272332
Average (Standard dev.)0.0014049211 (±0.04255815)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions320320320
Spacing320320320
CellA=B=C: 345.6 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: Half map A of the final 3D reconstruction

Fileemd_44259_half_map_1.map
AnnotationHalf map A of the final 3D reconstruction
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Half map B of the final 3D reconstruction

Fileemd_44259_half_map_2.map
AnnotationHalf map B of the final 3D reconstruction
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Transient receptor potential cation channel subfamily M member 8

EntireName: Transient receptor potential cation channel subfamily M member 8
Components
  • Complex: Transient receptor potential cation channel subfamily M member 8
    • Protein or peptide: Transient receptor potential cation channel subfamily M member 8
  • Ligand: 3-{7-(trifluoromethyl)-5-[2-(trifluoromethyl)phenyl]-1H-benzimidazol-2-yl}-1-oxa-2-azaspiro[4.5]dec-2-ene
  • Ligand: CHOLESTEROL HEMISUCCINATE
  • Ligand: nonyl(oxo)di(propan-2-yl)-lambda~5~-phosphane
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose

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Supramolecule #1: Transient receptor potential cation channel subfamily M member 8

SupramoleculeName: Transient receptor potential cation channel subfamily M member 8
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1
Source (natural)Organism: Mus musculus (house mouse)

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Macromolecule #1: Transient receptor potential cation channel subfamily M member 8

MacromoleculeName: Transient receptor potential cation channel subfamily M member 8
type: protein_or_peptide / ID: 1 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 131.548312 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MASFEGARLS MRSRRNGTMG STRTLYSSVS RSTDVSYSDS DLVNFIQANF KKRECVFFTR DSKAMENICK CGYAQSQHIE GTQINQNEK WNYKKHTKEF PTDAFGDIQF ETLGKKGKYL RLSCDTDSET LYELLTQHWH LKTPNLVISV TGGAKNFALK P RMRKIFSR ...String:
MASFEGARLS MRSRRNGTMG STRTLYSSVS RSTDVSYSDS DLVNFIQANF KKRECVFFTR DSKAMENICK CGYAQSQHIE GTQINQNEK WNYKKHTKEF PTDAFGDIQF ETLGKKGKYL RLSCDTDSET LYELLTQHWH LKTPNLVISV TGGAKNFALK P RMRKIFSR LIYIAQSKGA WILTGGTHYG LMKYIGEVVR DNTISRNSEE NIVAIGIAAW GMVSNRDTLI RSCDDEGHFS AQ YIMDDFT RDPLYILDNN HTHLLLVDNG CHGHPTVEAK LRNQLEKYIS ERTSQDSNYG GKIPIVCFAQ GGGRETLKAI NTS VKSKIP CVVVEGSGQI ADVIASLVEV EDVLTSSMVK EKLVRFLPRT VSRLPEEEIE SWIKWLKEIL ESSHLLTVIK MEEA GDEIV SNAISYALYK AFSTNEQDKD NWNGQLKLLL EWNQLDLASD EIFTNDRRWE SADLQEVMFT ALIKDRPKFV RLFLE NGLN LQKFLTNEVL TELFSTHFST LVYRNLQIAK NSYNDALLTF VWKLVANFRR SFWKEDRSSR EDLDVELHDA SLTTRH PLQ ALFIWAILQN KKELSKVIWE QTKGCTLAAL GASKLLKTLA KVKNDINAAG ESEELANEYE TRAVELFTEC YSNDEDL AE QLLVYSCEAW GGSNCLELAV EATDQHFIAQ PGVQNFLSKQ WYGEISRDTK NWKIILCLFI IPLVGCGLVS FRKKPIDK H KKLLWYYVAF FTSPFVVFSW NVVFYIAFLL LFAYVLLMDF HSVPHTPELI LYALVFVLFC DEVRQWYMNG VNYFTDLWN VMDTLGLFYF IAGIVFRLHS SNKSSLYSGR VIFCLDYIIF TLRLIHIFTV SRNLGPKIIM LQRMLIDVFF FLFLFAVWMV AFGVARQGI LRQNEQRWRW IFRSVIYEPY LAMFGQVPSD VDSTTYDFSH CTFSGNESKP LCVELDEHNL PRFPEWITIP L VCIYMLST NILLVNLLVA MFGYTVGIVQ ENNDQVWKFQ RYFLVQEYCN RLNIPFPFVV FAYFYMVVKK CFKCCCKEKN ME SNACCFR NEDNETLAWE GVMKENYLVK INTKANDNSE EMRHRFRQLD SKLNDLKSLL KEIANNIKSN SLEVLFQGPD YKD DDDKAH HHHHHHHHH

UniProtKB: Transient receptor potential cation channel subfamily M member 8

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Macromolecule #2: 3-{7-(trifluoromethyl)-5-[2-(trifluoromethyl)phenyl]-1H-benzimida...

MacromoleculeName: 3-{7-(trifluoromethyl)-5-[2-(trifluoromethyl)phenyl]-1H-benzimidazol-2-yl}-1-oxa-2-azaspiro[4.5]dec-2-ene
type: ligand / ID: 2 / Number of copies: 4 / Formula: T14
Molecular weightTheoretical: 467.407 Da
Chemical component information

ChemComp-T14:
3-{7-(trifluoromethyl)-5-[2-(trifluoromethyl)phenyl]-1H-benzimidazol-2-yl}-1-oxa-2-azaspiro[4.5]dec-2-ene

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Macromolecule #3: CHOLESTEROL HEMISUCCINATE

MacromoleculeName: CHOLESTEROL HEMISUCCINATE / type: ligand / ID: 3 / Number of copies: 12 / Formula: Y01
Molecular weightTheoretical: 486.726 Da
Chemical component information

ChemComp-Y01:
CHOLESTEROL HEMISUCCINATE

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Macromolecule #4: nonyl(oxo)di(propan-2-yl)-lambda~5~-phosphane

MacromoleculeName: nonyl(oxo)di(propan-2-yl)-lambda~5~-phosphane / type: ligand / ID: 4 / Number of copies: 4 / Formula: ULO
Molecular weightTheoretical: 260.396 Da
Chemical component information

ChemComp-ULO:
nonyl(oxo)di(propan-2-yl)-lambda~5~-phosphane

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Macromolecule #5: 2-acetamido-2-deoxy-beta-D-glucopyranose

MacromoleculeName: 2-acetamido-2-deoxy-beta-D-glucopyranose / type: ligand / ID: 5 / Number of copies: 4 / Formula: NAG
Molecular weightTheoretical: 221.208 Da
Chemical component information

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 8
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 2 / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 293.15 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Specialist opticsEnergy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number grids imaged: 1 / Number real images: 14226 / Average electron dose: 60.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.2 µm / Nominal defocus min: 0.7000000000000001 µm / Nominal magnification: 81000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 4913002
Startup modelType of model: EMDB MAP
EMDB ID:

7127


Details: The EM map for the previously reported apo TRPM8 structure (EMD-7127) was low-pass filtered to 30-angstrom and used as an initial model without a reference mask.
Final reconstructionApplied symmetry - Point group: C4 (4 fold cyclic) / Resolution.type: BY AUTHOR / Resolution: 2.76 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC / Number images used: 112062
Initial angle assignmentType: PROJECTION MATCHING / Software - Name: RELION
Final angle assignmentType: PROJECTION MATCHING / Software - Name: cryoSPARC
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelChain - Source name: Other / Chain - Initial model type: experimental model
Details: Another structural model from this study was used as the initial model for model building.
RefinementSpace: REAL / Protocol: RIGID BODY FIT
Output model

PDB-9b6h:
Cryo-EM structure of the mouse TRPM8 channel in complex with the antagonist TC-I 2014 and the cooling agonist C3

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