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Structure paper

TitleStructural and functional analyses of a GPCR-inhibited ion channel TRPM3.
Journal, issue, pagesNeuron, Vol. 111, Issue 1, Page 81-91.e7, Year 2023
Publish dateJan 4, 2023
AuthorsChen Zhao / Roderick MacKinnon /
PubMed AbstractG-protein coupled receptors (GPCRs) govern the physiological response to stimuli by modulating the activity of downstream effectors, including ion channels. TRPM3 is an ion channel inhibited by GPCRs ...G-protein coupled receptors (GPCRs) govern the physiological response to stimuli by modulating the activity of downstream effectors, including ion channels. TRPM3 is an ion channel inhibited by GPCRs through direct interaction with G protein (Gβγ) released upon their activation. This GPCR-TRPM3 signaling pathway contributes to the analgesic effect of morphine. Here, we characterized Gβγ inhibition of TRPM3 using electrophysiology and single particle cryo-electron microscopy (cryo-EM). From electrophysiology, we obtained a half inhibition constant (IC50) of ∼240 nM. Using cryo-EM, we determined structures of mouse TRPM3 expressed in human cells with and without Gβγ and with and without PIP, a lipid required for TRPM3 activity, at resolutions of 2.7-4.7 Å. Gβγ-TRPM3 interfaces vary depending on PIP occupancy; however, in all cases, Gβγ appears loosely attached to TRPM3. The IC50 in electrophysiology experiments raises the possibility that additional unknown factors may stabilize the TRPM3-Gβγ complex.
External linksNeuron / PubMed:36283409
MethodsEM (single particle)
Resolution2.7 - 4.7 Å
Structure data

EMDB-27338, PDB-8ddq:
cryo-EM structure of TRPM3 ion channel in the presence of soluble Gbg, focused on channel
Method: EM (single particle) / Resolution: 2.7 Å

EMDB-27339, PDB-8ddr:
cryo-EM structure of TRPM3 ion channel in the absence of PIP2
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-27340, PDB-8dds:
cryo-EM structure of TRPM3 ion channel in the presence of PIP2, state1
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-27341: Cryo-EM structure of TRPM3 ion channel in the presence of PIP2, state2
PDB-8ddt: cryo-EM structure of TRPM3 ion channel in the presence of PIP2, state2
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-27342, PDB-8ddu:
cryo-EM structure of TRPM3 ion channel in the presence of PIP2, state3
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-27343: cryo-EM structure of TRPM3 ion channel in the presence of PIP2, state4
PDB-8ddv: Cryo-EM structure of TRPM3 ion channel in the presence of PIP2, state4
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-27344, PDB-8ddw:
cryo-EM structure of TRPM3 ion channel in complex with Gbg, tethered by ALFA-nanobody
Method: EM (single particle) / Resolution: 4.7 Å

EMDB-27345, PDB-8ddx:
cryo-EM structure of TRPM3 ion channel in complex with Gbg in the presence of PIP2, tethered by ALFA-nanobody
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-28031, PDB-8ed7:
cryo-EM structure of TRPM3 ion channel in apo state
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-28032, PDB-8ed8:
cryo-EM structure of TRPM3 ion channel in the presence of PIP2 and PregS, state 1
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-28033, PDB-8ed9:
cryo-EM structure of TRPM3 ion channel in the presence with PIP2 and PregS, state 2
Method: EM (single particle) / Resolution: 3.4 Å

Chemicals

ChemComp-3PH:
1,2-DIACYL-GLYCEROL-3-SN-PHOSPHATE

ChemComp-9Z9:
(3beta,14beta,17beta,25R)-3-[4-methoxy-3-(methoxymethyl)butoxy]spirost-5-en / detergent*YM

ChemComp-NA:
Unknown entry

ChemComp-PIO:
[(2R)-2-octanoyloxy-3-[oxidanyl-[(1R,2R,3S,4R,5R,6S)-2,3,6-tris(oxidanyl)-4,5-diphosphonooxy-cyclohexyl]oxy-phosphoryl]oxy-propyl] octanoate

Source
  • mus musculus (house mouse)
  • homo sapiens (human)
KeywordsMEMBRANE PROTEIN / TRPM3 / ion channel / Gbg / PIP2

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