Structure of the Full-Length TRPV2 Channel by Cryo-EM

Huynh, Kevin W.; Cohen, Matthew R.; Jiang, Junchen; Samanta, Amrita; Lodowski, David T.; Zhou, Z. Hong; Moiseenkova-Bell, Vera Y.

doi:10.1017/s1431927616006437

Cited by 14 publications

(27 citation statements)

References 43 publications

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“…1C) (17). This finding confirmed the overall structural similarity between TRPV1 and TRPV2 (11,18,19), which would permit introduction of capsaicin binding to TRPV2. Rather than transferring the entire capsaicin-binding pocket as in the previous study (9), here, we aim to convey vanilloid sensitivity to TRPV2 with minimal structural perturbations by rationally mutating critical residues we identified to be essential for ligand gating of TRPV1 (6).…”

supporting

confidence: 77%

“…Conformational change in S4 is coupled to the movement of the S4-S5 linker to induce a conformational change in S6 interacting with the linker, leading to the opening of the activation gate (35)(36)(37). Although the S1-S4 domains of TRP channels are structurally similar to that of Kv channels (11,18,19,38,39), no positively charged residue exists in the S4 of TRPV channels so this domain is expected to remain stationary upon membrane depolarization or the presence of other activators (30). This allows capsaicin or resiniferatoxin to first secure itself to the stationary S1-S4 domain, and then stabilize the outward conformation of the S4-S5 linker to mediate S6 movements leading to channel activation (6,40).…”

Section: Discussionmentioning

confidence: 99%

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Rational design and validation of a vanilloid-sensitive TRPV2 ion channel

Yang

Yarov‐Yarovoy

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Vanilloids activation of TRPV1 represents an excellent model system of ligand-gated ion channels. Recent studies using cryo-electron microcopy (cryo-EM), computational analysis, and functional quantification revealed the location of capsaicin-binding site and critical residues mediating ligand-binding and channel activation. Based on these new findings, here we have successfully introduced high-affinity binding of capsaicin and resiniferatoxin to the vanilloid-insensitive TRPV2 channel, using a rationally designed minimal set of four point mutations (F467S-S498F-L505T-Q525E, termed TRPV2_Quad). We found that binding of resiniferatoxin activates TRPV2_Quad but the ligand-induced open state is relatively unstable, whereas binding of capsaicin to TRPV2_Quad antagonizes resiniferatoxin-induced activation likely through competition for the same binding sites. Using Rosettabased molecular docking, we observed a common structural mechanism underlying vanilloids activation of TRPV1 and TRPV2_Quad, where the ligand serves as molecular "glue" that bridges the S4-S5 linker to the S1-S4 domain to open these channels. Our analysis revealed that capsaicin failed to activate TRPV2_Quad likely due to structural constraints preventing such bridge formation. These results not only validate our current working model for capsaicin activation of TRPV1 but also should help guide the design of drug candidate compounds for this important pain sensor.on channels constitute the second largest family of drug targets for therapeutics (1-3); therefore, understanding their gating mechanisms by small-molecule ligands is critical for both basic and translational research. Capsaicin activation of the pain-sensing transient receptor potential vanilloid 1 (TRPV1) ion channel represents an outstanding model system for understanding liganddependent gating process (4), because capsaicin not only potently activates the channel with a submicromolar EC 50 (5) but also effectively stabilizes the channel at high open probability (6-8). Moreover, capsaicin activation is highly selective for TRPV1 channel (5). Previous mutagenesis (9, 10) and cryo-EM studies (11,12) have shown that capsaicin binds near the third (S3) and fourth (S4) transmembrane segments of TRPV1 (Fig. 1A). Based on the high-resolution cryo-EM structures, we have used a combination of computational and functional assays to reveal that capsaicin takes a "tail-up, head-down" configuration in the ligand-binding pocket (6) (Fig. 1B). The aliphatic tail forms extensive but nonspecific van der Waals (VDW) interactions with residues lining the binding pocket, whereas the vanillyl group (head) and the amide group (neck) of capsaicin form a hydrogen bond with E571 on S4-S5 linker and T551 on S4, respectively. To activate TRPV1, VDW interactions first secure capsaicin in the pocket and the neck of capsaicin forms a hydrogen bond with T551. This is followed by the formation of another hydrogen bond between the head and E571, which stabilizes the outward conformation of S4-S5 linker to open the chann...

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supporting

confidence: 77%

Section: Discussionmentioning

confidence: 99%

Rational design and validation of a vanilloid-sensitive TRPV2 ion channel

Yang

Yarov‐Yarovoy

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“… a–i , Pairwise superposition of the pore domain in hTRPV6 with ( a ) rat TRPV1 18 (PDB ID: 5IRX; RMSD = 2.065 Å), ( b ) rabbit TRPV2 21 (PDB ID: 5AN8; RMSD = 3.757 Å), ( c ) rat TRPV2 24 (PDB ID: 5HI9; RMSD = 4.399 Å), ( d ) human TRPA1 23 (PDB ID: 3J9P; RMSD = 1.429 Å), ( e ) human PKD2 25 (PDB ID: 5T4D; RMSD = 2.676 Å), ( f ) KcsA from Streptomyces lividans 47 (PDB ID: 1BL8; RMSD = 2.708 Å), ( g ) MthK from M. thermautotrophicum 48 (PDB ID: 1LNQ; RMSD = 2.947 Å), ( h ) rat Shaker 49 (PDB ID: 2A79; RMSD = 2.487 Å) and ( i ) rat GluA2 AMPA-subtype iGluR 28 (PDB ID: 5WEO; RMSD = 2.044 Å). j , Sequence alignment for the pore region of human TRPV3–6, TRPA1 and PKD2, rat TRPV1, 2, 6, Shaker and GluA2, rabbit TRPV2 and bacterial K + channels KcsA and MthK.…”

Section: Extended Datamentioning

confidence: 99%

Opening of the human epithelial calcium channel TRPV6

McGoldrick

Singh

Saotome

et al. 2017

Nature

189

322

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Ca2+-selective transient receptor potential vanilloid subfamily member 6 (TRPV6) channels play a critical role in calcium uptake in epithelial tissues1–4. Altered TRPV6 expression is associated with a variety of human diseases5, including cancers6. TRPV6 channels are constitutively active1,7,8 and their open probability depends on the lipidic composition of the membrane, increasing significantly in the presence of phosphatidylinositol 4,5-bisphosphate (PIP2)7,9. We previously solved crystal structures of detergent-solubilized rat TRPV6 in the closed state10,11. Corroborating previous electrophysiological studies3, these structures demonstrated that the Ca2+ selectivity of TRPV6 arises from a ring of aspartate side chains in the selectivity filter that tightly binds Ca2+. However, it has remained unknown how TRPV6 channels open and close their pores for ion permeation. Here we present cryo-EM structures of human TRPV6 in the open and closed states. The channel selectivity filter adopts similar conformations in both states, consistent with its explicit role in ion permeation. The iris-like channel opening is accompanied by an α-to-π helical transition in the pore-lining S6 helices at an alanine hinge just below the selectivity filter. As a result of this transition, the S6 helices bend and rotate, exposing different residues to the ion channel pore in the open and closed states. This novel gating mechanism, which defines the constitutive activity of TRPV6, is unique for tetrameric ion channels and provides new structural insights for understanding their diverse roles in physiology and disease.

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“…In silico molecular docking studies have been used to support their pharmacological results. To date, research regarding the effect of endogenous ligand on TRPV2 channels is scarce [38][39]. Most published studies were carried out using a TRPV1 model [40][41][42][43].…”

Section: Pocketmentioning

confidence: 99%

“…TRPV1 shares about 50 % sequence identity with TRPV2 [46]. Huynh et al recently reported the structure of the full-length rat TRPV2 channel by cryo-electron microscopy [38], while Zubcevic et al reported the atomic model of rabbit TRPV2 [39]. In crystallographic analyzes of human TRPV2, there is limited information on the structure of the ankyrin repeat domain of TRPV2.…”

Section: Pocketmentioning

confidence: 99%

Acute exposure to an electric field induces changes in human plasma lysophosphatidylcholine (lysoPC)-22:4 levels: Molecular insight into the docking of lysoPC-22:4 interaction with TRPV2

Nakagawa-Yagi¹,

Hara²,

Nakanishi³

et al. 2017

Integr Mol Med

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Medical treatment using high-voltage electric potential (HELP) devices to generate an electric field (EF) is an alternative therapy commonly used in Japan. However, the underlying mechanisms of the potential health benefits are not fully understood. To address this issue, we investigated the levels of lyso-form phospholipids using selected reaction monitoring (SRM) analysis in plasma samples obtained from healthy human subjects before and after a single HELP exposure (9 kV/electrode + 9 kV/electrode, 30 min). Lysophosphatidylcholine (lysoPC)-22:4 was significantly upregulated after HELP exposure. However, there was no effect on the levels of lysophosphatidic acid (lysoPA), or other lysoPC species. LysoPC is known to accelerate intestinal movement as a putative endogenous activator of transient receptor potential vanilloid 2 (TRPV2). We further examined the in silico docking simulation of lysoPC-22:4 with TRPV2. Docking results showed that lysoPC-22:4 has good binding energy (-8.2 kcal/mol). Our findings provide new insight into the molecular mechanisms of constipation alleviation by EF therapy.

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Structure of the Full-Length TRPV2 Channel by Cryo-EM

Cited by 14 publications

References 43 publications

Rational design and validation of a vanilloid-sensitive TRPV2 ion channel

Rational design and validation of a vanilloid-sensitive TRPV2 ion channel

Opening of the human epithelial calcium channel TRPV6

Acute exposure to an electric field induces changes in human plasma lysophosphatidylcholine (lysoPC)-22:4 levels: Molecular insight into the docking of lysoPC-22:4 interaction with TRPV2

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