TRPC channels: Structure, function, regulation and recent advances in small molecular probes

Wang, Hongbo; Cheng, Xiaoding; Tian, Jin Bin; Xiao, Yuling; Tian, Tian; Xu, Fuchun; Hong, Xuechuan; Zhu, Michael X.

doi:10.1016/j.pharmthera.2020.107497

Cited by 131 publications

(130 citation statements)

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“…The widely expressed TRPC1/4/5 channels have been implicated in various physiological and pathological mechanisms and are an emerging class of potential drug targets. 4,5,10,17 Our study provides the first structural insight into small-molecule modulation of a TRPC1/4/5 channel. By determination of a 3.0 Å structure of the TRPC5:C5 channel in complex with the TRPC1/4/5 inhibitor Pico145, we show that Pico145 can bind to up to four lipid binding sites between TRPC5 subunits, where it displaces phospholipids that interact with the channel's pore helices, previously proposed to be ceramide-1-phosphate or a phosphatidic acid.…”

Section: Discussionmentioning

confidence: 87%

“…Transient Receptor Potential Canonical (TRPC) proteins form homo-or heterotetrameric, nonselective cation channels permeable by Na + and Ca 2+ . [1][2][3][4][5][6] Based on sequence similarity, the mammalian TRPC proteins can be further divided into sub-groups: TRPC1/4/5, TRPC3/6/7 and TRPC2, 7 the latter of which is encoded by a pseudogene in humans. 8 TRPC4 and TRPC5 are the most closely related TRPC proteins 9 (70% sequence identity) and can form functional, homomeric TRPC4:C4 and TRPC5:C5 channels.…”

Section: Introductionmentioning

confidence: 99%

“…[10][11][12][13][14][15] Although disruption of the Trpc4/5 genes 16 and global expression of a dominant-negative mutant TRPC5 12 do not cause catastrophic phenotypes in rodents, TRPC1/4/5 channels have been implicated in a wide range of physiological and pathological mechanisms. 4,5,10,17 These findings have driven the development of potent and selective TRPC1/4/5 modulators as chemical probes and drug candidates, 5,10,18 and clinical trials have been started by Hydra Biosciences/Boehringer Ingelheim (the TRPC4/5 channel inhibitor BI 135889 for treatment of anxiety/CNS disorders) and Goldfinch Bio (the TRPC5 channel inhibitor GFB-887 for genetically-driven kidney disease).…”

Section: Introductionmentioning

confidence: 99%

“…Physiological activation and modulation of TRPC1/4/5 channel activity is complex, [4][5][6] and may include mediation by endogenous and dietary lipids. 12,[19][20][21][22] In addition, structurally diverse pharmacological modulators have been reported, 5,10,18 including inhibitors suitable for studies of TRPC1/4/5 in cells, tissues and animal models such as the xanthines Pico145 (also called HC-608) [23][24][25] and HC-070, 24,25 the pyridazinone derivative GFB-8438, 26 and the benzimidazole ML204. 27 However, structural insight into the mode-of-action of small-molecule TRPC1/4/5 modulators is lacking, and no small-molecule binding sites have been identified.…”

Section: Introductionmentioning

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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Section: Discussionmentioning

confidence: 87%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

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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“…Several available pharmacological tool compounds permit the identification of TRPC5 as putative drug targets for certain diseases ( 21-23 ). (-)-englerin A (EA), a natural product obtained from Phyllanthus engleri , can selectively activate TRPC4/5 and therefore inhibit tumor growth ( 24, 25 ), suggesting activation of TRPC4/5 might be a strategy to cure certain types of cancer.…”

Section: Introductionmentioning

confidence: 99%

Structural basis for human TRPC5 channel inhibition by two distinct inhibitors

Song

Wei

Guo

et al. 2020

Preprint

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17TRPC5 channel is a non-selective cation channel that participates diverse physiological 18 processes. Human TRPC5 inhibitors show promise in the treatment of anxiety disorder, 19 depression and kidney disease. Despite the high relevance of TRPC5 to human health, its 20 inhibitor binding pockets have not been fully characterized due to the lack of structural 21 information, which greatly hinders structure-based drug discovery. Here we show cryo-EM 22 structures of human TRPC5 in complex with two distinct inhibitors, namely clemizole and 23 HC-070, to the resolution of 2.7 Å. Based on the high-quality cryo-EM maps, we uncover the 24 different binding pockets and detailed binding modes for these two inhibitors. Clemizole 25 binds inside the voltage sensor-like domain of each subunit, while HC-070 binds close to the 26 ion channel pore and is wedged between adjacent subunits. Both of them exert the inhibitory 27 function by stabilizing the ion channel in a closed state. These structures provide templates 28 for further design and optimization of inhibitors targeting human TRPC5. 29 70 inhibitory mechanism of these two distinct inhibitors against human TRPC5 (hTRPC5), we 71 embarked structural studies of hTRPC5 in complex with CMZ or HC-070. Our high 72 resolution cryo-EM maps are sufficient to identify their binding pockets and will aid further 73 drug development. 74 75 76 Results 77 78 Structure of human TRPC5 in complex with inhibitors 79To reveal the binding sites of CMZ and HC-070 on hTRPC5, we expressed hTRPC5 in 80 HEK293F cells for structural studies. Similar to the mouse TRPC5 counterpart (mTRPC5) 81 (33), C-terminal truncation of hTRPC5 generated a construct (hTRPC5 1-764 ) that showed 82 higher expression level of the tetramer in comparison with the full length wild type hTRPC5 83 ( Fig. S1A). Moreover, we found hTRPC5 1-764 retained the pharmacological properties of full 84 length hTRPC5 such as activation by EA ( Fig. S1, B to E). Therefore, we used hTRPC5 1-764 85 protein for structural studies. Tetrameric hTRPC5 1-764 channel was purified in detergent 86 micelles (Fig. S1F and G). To obtain the CMZ-bound and the HC-070-bound TRPC5 87 structures, CMZ or HC-070 was added to the concentrated hTRPC5 1-764 protein for cryo-EM 88 sample preparation. We obtained maps of hTRPC5 in CMZ-bound state and HC-070-bound 89 state to the resolution of 2.7 Å (Fig. 1, Figs. S2 to S5; table S1). The map qualities were 90 sufficient for model building and ligand assignment (Fig. S3, D and E, and Fig. S5, D and E).91The overall architecture of inhibitor-bound hTRPC5 is similar to the structure of mTRPC5 in 92 apo state (33), occupying 100 Å × 100 Å × 130 Å in three-dimensional space (Fig. 1). The 93 four-fold symmetric channel has two layer architecture, composed of the intracellular 94 cytosolic domain (ICD) layer and the transmembrane domain (TMD) layer (Fig. 1). In ICD 95 layer, the N-terminal ankyrin repeats domain (ARD) is below the linker-helix domain (LHD) 96 region (Fig .1F). The TRP helix mediates interactions...

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The Calcium Signaling Mechanisms in Arterial Smooth Muscle and Endothelial Cells

Ottolini

Sonkusare

2021

Comprehensive Physiology

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TRPC channels: Structure, function, regulation and recent advances in small molecular probes

Cited by 131 publications

References 363 publications

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

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

Structural basis for human TRPC5 channel inhibition by two distinct inhibitors

The Calcium Signaling Mechanisms in Arterial Smooth Muscle and Endothelial Cells

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