Structure of the human lipid-gated cation channel TRPC3

Chen, Fan; Choi, Woo-Young; Sun, Weinan; Du, Juan; Lü, Wei

doi:10.7554/elife.36852

Cited by 115 publications

(154 citation statements)

References 58 publications

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“…2C and Table 1). Finally, only TRPV2, TRPV4, TRPV5, TRPM8 and TRPC3 do not show a π-helical segment in S6 (31)(32)(33)(34)(35)(36)(37). Either these TRP channels lack altogether the π-helical conformation or, alternatively, this conformation has not been experimentally captured yet.…”

Section: Resultsmentioning

confidence: 90%

A consistent picture of TRPV1 activation emerges from molecular simulations and experiments

Yudin

et al. 2018

Preprint

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Although the structure of TRPV1 has been experimentally determined in both the closed and open states, very little is known about its activation mechanism. In particular, the conformational changes occurring in the pore domain and resulting in ionic conduction have not been identified yet. Here, we suggest a hypothetical molecular mechanism for TRPV1 activation, which involves the rotation of a conserved asparagine in S6 from the S4-S5 linker toward the pore. This rotation is correlated with the dehydration of four peripheral cavities located between S6 and the S4-S5 linker and the hydration of the pore. In light of our hypothesis, we perform bioinformatics analyses of TRP and other evolutionary related ion channels, analyze newly available structures and re-examine previously reported water accessibility and mutagenesis experiments. Overall, we provide several independent lines of evidence that corroborate our hypothesis. Finally, we show that the proposed molecular mechanism is compatible with the currently existing idea that in TRPV1 the selectivity filter acts as a secondary gate.

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Section: Resultsmentioning

confidence: 90%

A consistent picture of TRPV1 activation emerges from molecular simulations and experiments

Yudin

et al. 2018

Preprint

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“…The TRPC5 lipid binding site that Pico145 interacts with is highly conserved within the TRPC family (Supplementary Table 2). Bound (phospho)lipids have been found in this site in structures of hTRPC3, 35,37 mTRPC4, 33 mTRPC5, 34 and hTRPC6, 35,36 and the TRPC6 activator AM-0883 displaces the ordered lipid bound in this site near the P-loop of hTRPC6 (Supplementary Figure 5F). 36 The hTRPC5 residues F576 and W577 are conserved throughout the TRPC family and are likely to be responsible for lipid binding.…”

Section: Discussionmentioning

confidence: 97%

“…Indeed, a similar density is present in the mTRPC4 structure ( Figure 2C). The hTRPC3 35,37 and hTRPC6 35,36 structures contain lipids that interact with their LFW motifs as well, but with altered geometry (Supplementary Figure 5). Our TRPC5:Pico145 structure also showed density in this region.…”

Section: Pico145 Binds To a Conserved Lipid Binding Site Of Trpc5mentioning

confidence: 99%

“…In the zinc binding domain, the densities were similar (and partially disordered) in the TRPC5:Zn 2+ and TRPC5:Pico145 structures, suggesting that the site may already be fully occupied in the absence of added ZnCl2. A structural alignment showed that the putative zinc binding motif is present in the electron density maps of all TRPC channel structures; it was built into the hTRPC3 35,37 and hTRPC6 35,36 structures, but not into the mTRPC4 33 or mTRPC5 34 structures (Supplementary Figure 10). These findings suggest that TRPC channels have a conserved intracellular zinc binding site.…”

Section: Mutations In the Xanthine Binding Site Alter The Response Ofmentioning

confidence: 99%

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Cryo-EM structures of human TRPC5 reveal interaction of a xanthine-based TRPC1/4/5 inhibitor with a conserved lipid binding site

Wright

Simmons

et al. 2020

Preprint

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TRPC1/4/5 channels are non-specific cation channels implicated in a wide variety of diseases, and TRPC1/4/5 inhibitors have recently entered the first clinical trials. However, fundamental and translational studies require a better understanding of TRPC1/4/5 channel regulation by endogenous and exogenous factors. Although several potent and selective TRPC1/4/5 modulators have been reported, the paucity of mechanistic insights into their modes-of-action remains a barrier to the development of new chemical probes and drug candidates. The xanthine class of modulators includes the most potent and selective TRPC1/4/5 inhibitors described to date, as well as TRPC5 activators. Our previous studies suggest that xanthines interact with a, so far, elusive pocket of TRPC1/4/5 channels that is essential to channel gating. Targeting this pocket may be a promising strategy for TRPC1/4/5 drug discovery. Here we report the first structure of a small molecule-bound TRPC1/4/5 channelhuman TRPC5 in complex with the xanthine Pico145to 3.0 Å. We found that Pico145 binds to a conserved lipid binding site of TRPC5, where it displaces a bound phospholipid. Our findings explain the mode-of-action of xanthine-based TRPC1/4/5 modulators, and suggest a structural basis for TRPC1/4/5 modulation by endogenous factors such as (phospho)lipids and Zn 2+ ions. These studies lay the foundations for the structure-based design of new generations of TRPC1/4/5 modulators.

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“…The cryo-EM revolution has enabled recent structural solutions for TRPC3, TRPC4, and TRPC6 12-14 . Here we present the structure of the mouse TRPC5 at pH 7.5 to an overall resolution of 2.9 Å.…”

Section: Introductionmentioning

confidence: 99%

Cryo-EM structure of the receptor-activated TRPC5 ion channel at 2.9 angstrom resolution

Duan

Chen

et al. 2018

Preprint

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32The transient receptor potential canonical subfamily member 5 (TRPC5) is a 33 non-selective calcium-permeant cation channel. As a depolarizing channel, its function 34 is studied in the central nervous system and kidney. TRPC5 forms heteromultimers 35 with TRPC1, but also forms homomultimers. It can be activated by reducing agents 36 through reduction of the extracellular disulfide bond. Here we present the 2.9 Å 37 resolution electron cryo-microscopy (cryo-EM) structure of TRPC5. The structure of 38 TRPC5 in its apo state is partially open, which may be related to the weak activation of 39 TRPC5 in response to extracellular pH. We also report the conserved negatively charged 40 residues of the cation binding site located in the hydrophilic pocket between S2 and S3. 41Comparison of the TRPC5 structure to previously determined structures of other TRPC 42 and TRP channels reveals differences in the extracellular pore domain and in the 43 length of the S3 helix. Together, these results shed light on the structural features that 44 contribute to the specific activation mechanism of the receptor-activated TRPC5. 45 46 47 48 65 Results 66Overall structure of the mouse TRPC5 tetrameric ion channel 67 A mouse TRPC5 construct lacking the C-terminal 210 residues (a.a. 1-765, excluding a.a. 68 766-975) yielded more protein after purification than that of the full-length construct. The 69 baculovirus construct, consisting of a maltose binding protein (MBP) tag at the N 70 terminus, was stably purified to homogeneity in n-dodecyl β-D-maltoside (DDM) and 71 reconstituted in the amphipol poly (maleic anhydride-alt-1-decene) substituted with 72 3-(dimethylamino) propylamine (PMAL-C8; Supplementary Fig. 1). The amphipol 73 reconstituted detergent-free protein was then negatively stained and analyzed by 74 single-particle cryo-EM. 75Single particle cryo-EM analyses of TRPC5 at an overall resolution of ~ 2.9 Å was 76 sufficient for de novo model building (Supplementary Fig. 2, Supplementary Fig. 3, 77 Supplementary Table 1). Disordered regions led to poor densities for 7 residues in the 78 S1-S2 loop, 28 residues in the distal N terminus, and 3 residues in the truncated distal C 79 terminus. Similar to other solved TRP channel structures, TRPC5 forms a four-fold 80 symmetric homotetramer (Fig. 1a, b) with dimensions of 100 Å × 100 Å × 120 Å. 81Each of the four monomers can be divided into a compact cytosolic domain and a 82 transmembrane domain (TMD) (Fig. 1). The cytosolic domain is composed of the 83 N-terminal region with an ankyrin repeats domain (ARD) and seven α-helices (HLH); 84 the C-terminal subdomain contains a connecting helix and a coiled-coil domain. The 85 transmembrane domain (TMD) is composed of six α-helices (S1-S6), a TRP domain, and 315We thank Dr. Steve Harrison and the Cryo-EM Facility (Harvard Medical School) for use 316 of their microscopes. We thank Dr. Maofu Liao for providing the Python scripts and help in 317

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Structure of the human lipid-gated cation channel TRPC3

Cited by 115 publications

References 58 publications

A consistent picture of TRPV1 activation emerges from molecular simulations and experiments

A consistent picture of TRPV1 activation emerges from molecular simulations and experiments

Cryo-EM structures of human TRPC5 reveal interaction of a xanthine-based TRPC1/4/5 inhibitor with a conserved lipid binding site

Cryo-EM structure of the receptor-activated TRPC5 ion channel at 2.9 angstrom resolution

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