Structure of the cold- and menthol-sensing ion channel TRPM8

Yin, Ying; Wu, Mengyu; Zubcevic, Lejla; Borschel, William F.; Lander, Gabriel C.; Lee, Seok‐Yong

doi:10.1126/science.aan4325

Cited by 242 publications

(326 citation statements)

References 51 publications

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“…1c). Similar to the previously reported TRPM2, TRPM4, and TRPM8 structures 8,[26][27][28][29][30][31] , each TRPM2 DR protomer contains an N-terminal region composed of MHR1 to MHR4, a transmembrane channel region, and a C-terminal region. The CC2 serves as a link between the C-terminal NUDT9H domain and the rest of the channel (Fig.…”

Section: Structure Determination and Overall Architecture Of Trpm2 Drsupporting

confidence: 82%

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Visualizing structural transitions of ligand-dependent gating of the TRPM2 channel

Yin

Hsu

et al. 2019

Preprint

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The calcium-permeable transient receptor potential melastatin 2 (TRPM2) channel plays a key role in redox sensation in many cell types 1-3 . Channel activation requires binding of both ADP-ribose (ADPR) 2,4-6 and Ca 2+ 7 . The recently published TRPM2 structures from Danio rerio in the ligand-free and in the ADPR/Ca 2+ -bound conditions represent the channel in closed and open states, which uncover substantial tertiary and quaternary conformational rearrangements 8 .However, it is unclear how these rearrangements occur within the tetrameric channel during channel gating. Here we report two cryo-electron microscopy structures of TRPM2 from the same species in complex with Ca 2+ alone, and with both ADPR and Ca 2+ , determined to an overall resolution of ~3.8 Å and ~4.2 Å respectively. In comparison with the published results, our studies capture TRPM2 in two-fold symmetric intermediate states, offering a glimpse of the structural transitions within the tetramer that bridge the closed and open conformations.3 MainThe transient receptor potential melastatin (TRPM) ion channel family is part of the TRP channel superfamily and comprises eight members (TRPM1 to TRPM8) that carry out diverse functions in a variety of physiological pathways 9 . TRPM2 is a calcium-permeable non-selective cation channel that is widely expressed in the nervous, immune, and endocrine systems, and plays a crucial role in warmth and redox-dependent signaling 1-3 . Studies of TRPM2-deficient mice have shown that TRPM2 channels expressed in sensory and central neurons are responsible for sensation of warm temperatures and body temperature regulation 10,11 . TRPM2 has also been found to be activated by reactive oxygen species (ROS), consequently playing a key role in Ca 2+ signaling involved in chemokine production, insulin production, and cell death under oxidative stress [12][13][14][15][16] .Extensive electrophysiological studies have shown that activation of TRPM2 by ROS is caused by an increase of intracellular ADP-ribose (ADPR), a TRPM2 agonist, and that both ADPR and Ca 2+ are required for channel activation 2,[4][5][6][7]17 . Notably, TRPM2 contains a putative enzyme domain, called the Nudix Hydrolase 9 Homology (NUDT9H) domain located at the C-terminus, which exhibits a high degree of homology to the mitochondrial ADP-ribose pyrophosphatase NUDT9 18,19 . As such, TRPM2 was initially classified as a channel-enzyme in which enzymatic activity was believed to be coupled to channel gating 2,18 . However, subsequent studies have since repudiated this mechanism, as the NUDT9H domain lacks enzymatic activity. Instead it has been suggested that the NUDT9H domain merely serves as a binding site for ADPR 20 .Recently, structures of the zebrafish Danio rerio TRPM2 were reported in the ligand-free closed state and in the ADPR/Ca 2+ -bound open state (hereafter referred to as TRPM2 DR_closed and TRPM2 DR_open respectively) 8 . This study not only revealed the location of the NUDT9H domain, but also showed that the Ca 2+ ion binds in the cavity formed...

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Section: Structure Determination and Overall Architecture Of Trpm2 Drsupporting

confidence: 82%

“…6a-c). While the TMD and two membraneproximal CD layers resemble the architecture observed in the TRPM8 and TRPM4 structures [28][29][30][31] , the additional second coiled coil (CC2) and the NUDT9H domain together comprise a unique bottom CD layer in the TRPM2 structures ( Fig. 1c).…”

Section: Structure Determination and Overall Architecture Of Trpm2 Drmentioning

confidence: 81%

Visualizing structural transitions of ligand-dependent gating of the TRPM2 channel

Yin

Hsu

et al. 2019

Preprint

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“…The tyrosine residues potentially involved in such modifications could be located in the C-terminal region, given that this region was reported to be implicated in the cold-induced gating of TRPM8 (Brauchi, Orta, Salazar, Rosenmann, & Latorre, 2006). Other putative tyrosine residues involved in the modulation of TRPM8 channel activity could be the conserved residues Tyr 826 and Tyr 836 situated on the S4 transmembrane helix, according to the recently published TRPM8 structure (Yin et al, 2018). Mutations neutralizing charged amino acids within the S4 helix and the S4-S5 linker region reduced the gating charge of TRPM8 (Voets, Owsianik, Janssens, Talavera, & Nilius, 2007).…”

Section: Discussionmentioning

confidence: 99%

Regulation of TRPM8 channel activity by Src‐mediated tyrosine phosphorylation

Manolache

Şelescu

Maier

et al. 2019

Journal Cellular Physiology

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The transient receptor potential melastatin type 8 (TRPM8) receptor channel is expressed in primary afferent neurons where it is the main transducer of innocuous cold temperatures and also in a variety of tumors, where it is involved in progression and metastasis. Modulation of this channel by intracellular signaling pathways has therefore important clinical implications. We investigated the modulation of recombinant and natively expressed TRPM8 by the Src kinase, which is known to be involved in cancer pathophysiology and inflammation. Human TRPM8 expressed in HEK293T cells is constitutively tyrosine phosphorylated by Src which is expressed either heterologously or endogenously. Src action on TRPM8 potentiates its activity, as treatment with PP2, a selective Src kinase inhibitor, reduces both TRPM8 tyrosine phosphorylation and cold‐induced channel activation. RNA interference directed against the Src kinase diminished the extent of PP2‐induced functional downregulation of TRPM8, confirming that PP2 acts mainly through Src inhibition. Finally, the effect of PP2 on TRPM8 cold activation was reproduced in cultured rat dorsal root ganglion neurons, and this action was antagonized by the protein tyrosine phosphatase inhibitor pervanadate, confirming that TRPM8 activity is sensitive to the cellular balance between tyrosine kinases and phosphatases. This positive modulation of TRPM8 by Src kinase may be relevant for inflammatory pain and cancer signaling.

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“…CH2 helices form a vertical four-helix bundle that further tightens the tetramer (Fig. 4g), reminiscent of TRPA1 and TRPM C-terminal four-helix bundle structure [28][29][30][31][32] . Uniquely, the four CH1-CH2 connection linkers pack tightly to form a knot-like structure on the top of CH2 ( Fig.…”

Section: Inter-subunit Interactions Mediate Htrpc6 Tetramer Assemblymentioning

confidence: 99%

“…These protrusions might harbor the extracellular calcium sites that are important for the channel activity regulation 43 . Among all of the available TRP channel structures, the TMD of hTRPC6 mostly resembles that of NOMPC 39 and TRPM [29][30][31][32] , as they share common structural elements of the pre-S1 elbow and the TRP re-entrant located after the TRP helix. According to the sequence alignments, these structural elements are also preserved in the drosophila TRP channel, and other mammalian TRPC channels (Supplementary information, Figure S4).…”

Section: Transmembrane Domain Of the Htrpc6 Channelmentioning

confidence: 99%

Structure of the receptor-activated human TRPC6 ion channel

Tang

Guo

Zheng³

et al. 2018

Preprint

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Abstract：TRPC6 is a receptor-activated nonselective cation channel that belongs to the family of canonical transient receptor potential (TRPC) channels. It is activated by diacylglycerol, a lipid second messenger. TRPC6 is involved in many physiological processes and implicated in human genetic diseases. Here we present the structure of human TRPC6 homotetramer in complex with a newly identified high affinity inhibitor BTDM solved by single-particle cryo-electron microscopy to 3.8 Å resolution. The structure shows a two-layer architecture, in which the bellshaped cytosolic layer holds the transmembrane layer. Extensive inter-subunit interactions of cytosolic domain, including N terminal ankyrin repeats and C terminal coiled-coil, contribute to the tetramer assembly. The high affinity inhibitor BTDM wedges between S5-S6 pore domain and voltage sensor-like domain to inhibit channel opening. Our structure uncovers the molecular architecture of TRPC channels and provides a structural basis for understanding the mechanism of these channels .All rights reserved. No reuse allowed without permission.(which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.

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Structure of the cold- and menthol-sensing ion channel TRPM8

Cited by 242 publications

References 51 publications

Visualizing structural transitions of ligand-dependent gating of the TRPM2 channel

Visualizing structural transitions of ligand-dependent gating of the TRPM2 channel

Regulation of TRPM8 channel activity by Src‐mediated tyrosine phosphorylation

Structure of the receptor-activated human TRPC6 ion channel

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