A Gate Hinge Controls the Epithelial Calcium Channel TRPV5

Wijst, Jenny van der; Leunissen, E. H. P.; Blanchard, Maxime; Venselaar, Hanka; Verkaart, Sjoerd; Paulsen, Candice E.; Bindels, René J. M.; Hoenderop, Joost G. J.

doi:10.1038/srep45489

Cited by 25 publications

(25 citation statements)

References 54 publications

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“…5c,d). Our structural observation is consistent with a recent mutagenesis study 7 that suggested a similar gating mechanism for the TRPV5 channel and implicated His582 in this mechanism.…”

Section: Resultssupporting

confidence: 92%

“…6). Finally, consistent with the recent mutagenesis and structural studies 7 , we determined that in addition to the Met578, the rotation of the Phe574 and possibly His582 into the pore may further block Ca 2+ ion flow through TRPV5 (Fig. 6).…”

Section: Discussionsupporting

confidence: 89%

“…In the absence of modulators, TRPV5 and TRPV6 are constitutively active channels 3 . Several endogenous modulators such as calmodulin and phosphatidylinositol 4,5-bisphosphate (PIP 2 ) have been found to stabilize these channels in the closed or open conformation, respectively 7,8 . Additionally, the small-molecule antifungals econazole and miconazole, which are commonly used topically to treat skin infections, have been shown to effectively inhibit both TRPV5 and TRPV6 (refs.…”

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confidence: 99%

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Structural basis of TRPV5 channel inhibition by econazole revealed by cryo-EM

Hughes

Lodowski

Huynh

et al. 2018

Nat Struct Mol Biol

121

143

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The transient receptor potential vanilloid 5 (TRPV5) channel is a member of the transient receptor potential (TRP) channel family, which is highly selective for Ca2+, that is present primarily at the apical membrane of distal tubule epithelial cells in the kidney and plays a key role in Ca2+ reabsorption. Here we present the structure of the full-length rabbit TRPV5 channel as determined using cryo-EM in complex with its inhibitor econazole. This structure reveals that econazole resides in a hydrophobic pocket analogous to that occupied by phosphatidylinositides and vanilloids in TRPV1, thus suggesting conserved mechanisms for ligand recognition and lipid binding among TRPV channels. The econazole-bound TRPV5 structure adopts a closed conformation with a distinct lower gate that occludes Ca2+ permeation through the channel. Structural comparisons between TRPV5 and other TRPV channels, complemented with molecular dynamics (MD) simulations of the econazole-bound TRPV5 structure, allowed us to gain mechanistic insight into TRPV5 channel inhibition by small molecules.

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“…5c,d). Our structural observation is consistent with a recent mutagenesis study 7 that suggested a similar gating mechanism for the TRPV5 channel and implicated His582 in this mechanism.…”

Section: Resultssupporting

confidence: 92%

Section: Discussionsupporting

confidence: 89%

mentioning

confidence: 99%

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Structural basis of TRPV5 channel inhibition by econazole revealed by cryo-EM

Hughes

Lodowski

Huynh

et al. 2018

Nat Struct Mol Biol

121

143

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“…To explore the influence of W583, we determined a structure of full length TRPV5 with a W583A point mutation in nanodisc to 2.85 Å resolution. Consistent with previously reported 'gain-of-function' phenotype for the W583 mutants (18), the lower gate of the TRPV5 W583A structure has an open conformation (Supplemental Fig. 3).…”

Section: Ion Permeation Pathwaysupporting

confidence: 89%

Structural insight into TRPV5 channel function and modulation

Dang

Goor

Wang

et al. 2018

Preprint

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TRPV5 (transient receptor potential vanilloid) is a unique calcium-selective TRP channel that is essential for calcium homeostasis. TRPV5 and its close homologue TRPV6 do not exhibit thermosensitivity or ligand-dependent activation, unlike other TRPV channels, but are constitutively opened at physiological membrane potentials. Here, we report high resolution electron cryo-microscopy (cryo-EM) structures of truncated and full length TRPV5 in lipid nanodisc, as well as a TRPV5 W583A mutant structure and a complex structure of TRPV5 with calmodulin (CaM). These structures highlight and explain functional differences between the thermosensitive and calcium-selective TRPV channels. An extended S1-S2 linker folds on top of the channel that might shield it from modulation by extracellular factors. Resident lipid densities in the homologous vanilloid pocket are different from those previously found in TRPV1, supporting a comparatively more rigid architecture of TRPV5. A ring of tryptophan residues (W583) at the bottom of the pore coordinates a density and mutation of W583 resultes in opening of the lower gate. Moreover, we provide structural insight into the calcium-dependent channel inactivation and propose a flexible stoichiometry for TRPV5 and CaM binding.

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“…2e). Indeed, W583 is conserved in TRPV6 and TRPV5 channels and is involved in regulation of calcium uptake, as evidenced by mutation W583A in TRPV5, which induces cell death due to increased calcium influx 19 . The density at the S6 helices bundle crossing, which was not observed in the pore of the closed-state rTRPV6 10,11 , most likely represents another permeant ion bound in the open pore of hTRPV6.…”

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confidence: 99%

Opening of the human epithelial calcium channel TRPV6

McGoldrick

Singh

Saotome

et al. 2017

Nature

194

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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A Gate Hinge Controls the Epithelial Calcium Channel TRPV5

Cited by 25 publications

References 54 publications

Structural basis of TRPV5 channel inhibition by econazole revealed by cryo-EM

Structural basis of TRPV5 channel inhibition by econazole revealed by cryo-EM

Structural insight into TRPV5 channel function and modulation

Opening of the human epithelial calcium channel TRPV6

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