Structural basis for human TRPC5 channel inhibition by two distinct inhibitors

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Abstract

TRPC5 channel is a non-selective cation channel that participates diverse physiological processes. TRPC5 inhibitors show promise in the treatment of anxiety disorder, depression and kidney disease. However, the binding sites and inhibitory mechanism of TRPC5 inhibitors remain elusive. Here we present the cryo-EM structures of human TRPC5 in complex with two distinct inhibitors, namely clemizole and HC-070, to the resolution of 2.7 Å. The structures reveal that clemizole binds inside the voltage sensor-like domain of each subunit. In contrast, HC-070 is wedged between adjacent subunits and replaces the glycerol group of a putative DAG molecule near the extracellular side. Moreover, we found mutations in the inhibitor binding pockets altered the potency of inhibitors. These structures suggest that both clemizole and HC-070 exert the inhibitory functions by stabilizing the ion channel in a non-conductive closed state. These results pave the way for further design and optimization of inhibitors targeting human TRPC5.

Data availability

The density maps of hTRPC5 have been deposited to the Electron Microscopy Data Bank (EMDB) under the accession number: EMD-30987 for apo hTRPC5, EMD-30575 for CMZ-bound hTRPC5 and EMD-30576 for HC-070-bound hTRPC5. Coordinates of atomic model have been deposited in the Protein Data Bank (PDB) under the accession number: 7E4T for apo hTRPC5, 7D4P for CMZ-bound hTRPC5 and 7D4Q for HC-070-bound hTRPC5.

The following data sets were generated

1. Miao Wei
2. Kangcheng Song and Lei Chen
(2020) cryo-EM structure of hTRPC5 in complex with Clemizole
EMDB, EMD-30575.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-30575
1. Miao Wei
2. Kangcheng Song and Lei Chen
(2020) cryo-EM structure of hTRPC5 in complex with Clemizole
PDB, 7D4P.

https://www.rcsb.org/structure/7D4P
1. Miao Wei
2. Kangcheng Song and Lei Chen
(2020) cryo-EM structure of hTRPC5 in complex with HC-070
EMDB, EMD-30576.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-30576
1. Miao Wei
2. Kangcheng Song and Lei Chen
(2020) cryo-EM structure of hTRPC5 in complex with HC-070
PDB, 7D4Q.

https://www.rcsb.org/structure/7D4Q
1. Miao Wei
2. Kangcheng Song and Lei Chen
(2020) cryo-EM structure of hTRPC5 in apo state
EMDB, EMD-30987.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-30987
1. Miao Wei
2. Kangcheng Song and Lei Chen
(2020) cryo-EM structure of hTRPC5 in apo state
PDB, 7E4T.

https://www.rcsb.org/structure/7E4T

The following previously published data sets were used

1. Tang Q
2. Guo W
3. Zheng L
4. Wu JX
5. Liu M
6. Zhou X
7. Zhang X
8. Chen L
(2018) Cryo-EM structure of human TRPC6 at 3.8A resolution
EMDB, EMD-6856.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-6856

Article and author information

Author details

Kangcheng Song

Institute of Molecular Medicine, Peking University, Beijing, China

Competing interests
The authors declare that no competing interests exist.
Miao Wei

Institute of Molecular Medicine, Peking University, Beijing, China

Competing interests
The authors declare that no competing interests exist.
Wenjun Guo

Institute of Molecular Medicine, Peking University, Beijing, China

Competing interests
The authors declare that no competing interests exist.
Li Quan

Institute of Molecular Medicine, Peking University, Beijjing, China

Competing interests
The authors declare that no competing interests exist.
Yunlu Kang

Institute of Molecular Medicine, Peking University, Beijing, China

Competing interests
The authors declare that no competing interests exist.
Jing-Xiang Wu

Institute of Molecular Medicine, Peking University, Beijing, China

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-9851-0065
Lei Chen

Institute of Molecular Medicine, Peking University, Beijjing, China

For correspondence
chenlei2016@pku.edu.cn

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-7619-8311

Funding

National Key Research and Development Program of China (2016YFA0502004)

Lei Chen

National Natural Science Foundation of China (91957201,31870833,31821091)

Lei Chen

National Natural Science Foundation of China (31900859)

Jing-Xiang Wu

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.