Structural basis for pharmacological modulation of the TRPC6 channel

Abstract
Data availability
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Abstract

Transient receptor potential canonical (TRPC) proteins form nonselective cation channels that play physiological roles in a wide variety of cells. Despite growing evidence supporting the therapeutic potential of TRPC6 inhibition in treating pathological cardiac and renal conditions, mechanistic understanding of TRPC6 function and modulation remains obscure. Here we report cryo-EM structures of TRPC6 in both antagonist-bound and agonist-bound states. The structures reveal two novel recognition sites for the small-molecule modulators corroborated by mutagenesis data. The antagonist binds to a cytoplasm-facing pocket formed by S1-S4 and the TRP helix, whereas the agonist wedges at the subunit interface between S6 and the pore helix. Conformational changes upon ligand binding illuminate a mechanistic rationale for understanding TRPC6 modulation. Furthermore, structural and mutagenesis analyses suggest several disease-related mutations enhance channel activity by disrupting interfacial interactions. Our results provide principles of drug action that may facilitate future design of small molecules to ameliorate TRPC6-mediated diseases.

Data availability

The low pass filtered and amplitude modified 3D cryo-EM density maps for TRPC6 in complex with antagonist AM-1473 (accession code: EMD-20954) and agonist AM-0883 (accession code: EMD-20953) have been deposited in the electron microscopy data bank. Atomic coordinates for TRPC6 in complex with antagonist AM-1473 (accession code: 6UZA) and agonist AM-0883 (accession code: 6UZ8) have been deposited in the protein data bank.

The following data sets were generated

1. Bai Y
2. Yu X
3. Huang X
4. Chen H
(2019) Cryo-EM structure of human TRPC6 in complex with antagonist AM-1473
RCSB Protein Data Bank, 6UZA.

http://www.rcsb.org/structure/6UZA
1. Bai Y
2. Yu X
3. Huang X
4. Chen H
(2019) Cryo-EM structure of human TRPC6 in complex with antagonist AM-1473
Electron Microscopy Data Bank, EMD-20954.

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-20954
1. Bai Y
2. Yu X
3. Huang X
4. Chen H
(2019) Cryo-EM structure of human TRPC6 in complex with agonist AM-0833
RCSB Protein Data Bank, 6UZ8.

http://www.rcsb.org/structure/6UZ8
1. Bai Y
2. Yu X
3. Huang X
4. Chen H
(2019) Cryo-EM structure of human TRPC6 in complex with agonist AM-0833
Electron Microscopy Data Bank, EMD-20953.

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-20953

Article and author information

Author details

Yonghong Bai

Molecular Engineering, Amgen Inc, Cambridge, United States

For correspondence
ybai80@gmail.com

Competing interests
Yonghong Bai, At the time of the study YB was affiliated with Amgen Research, Amgen Inc. and has no financial interests to declare. The author has no other competing interests to declare..

"This ORCID iD identifies the author of this article:" 0000-0002-4334-0916
Xinchao Yu

Molecular Engineering, Amgen Inc, South San Francisco, United States

For correspondence
xyu01@amgen.com

Competing interests
Xinchao Yu, XY is affiliated with Amgen Research, Amgen Inc. and has no financial interests to declare. The author has no other competing interests to declare..
Hao Chen

Protein Technologies, Amgen Inc, Cambridge, United States

Competing interests
Hao Chen, At the time of the study HC was affiliated with Amgen Research, Amgen Inc. and has no financial interests to declare. The author has no other competing interests to declare..
Daniel Horne

Medicinal Chemistry, Amgen Inc, Cambridge, United States

Competing interests
Daniel Horne, At the time of the study DH was affiliated with Amgen Research, Amgen Inc. and has no financial interests to declare. The author has no other competing interests to declare..
Ryan White

Medicinal Chemistry, Amgen Inc, Cambridge, United States

Competing interests
Ryan White, At the time of the study RW was affiliated with Amgen Research, Amgen Inc. and has no financial interests to declare. The author has no other competing interests to declare..
Xiaosu Wu

Cardiometabolic Disorders, Amgen Inc, South San Francisco, United States

Competing interests
Xiaosu Wu, XW is affiliated with Amgen Research, Amgen Inc. and has no financial interests to declare. The author has no other competing interests to declare..
Paul Lee

Discovery Technologies, Amgen Inc, Thousand Oaks, United States

Competing interests
Paul Lee, At the time of the study PL was affiliated with Amgen Research, Amgen Inc. and has no financial interests to declare. The author has no other competing interests to declare..
Yan Gu

Protein Technologies, Amgen Inc, Cambridge, United States

Competing interests
Yan Gu, At the time of the study YG was affiliated with Amgen Research, Amgen Inc. and has no financial interests to declare. The author has no other competing interests to declare..
Sudipa Ghimire-Rijal

Molecular Engineering, Amgen Inc, Cambridge, United States

Competing interests
Sudipa Ghimire-Rijal, SGR is affiliated with Amgen Research, Amgen Inc. and has no financial interests to declare. The author has no other competing interests to declare..
Daniel C-H Lin

Cardiometabolic Disorders, Amgen Inc, South San Francisco, United States

For correspondence
dclin@amgen.com

Competing interests
Daniel C-H Lin, DCHL is affiliated with Amgen Research, Amgen Inc. and has no financial interests to declare. The author has no other competing interests to declare..
Xin Huang

Molecular Engineering, Amgen Inc, Cambridge, United States

For correspondence
hxin@amgen.com

Competing interests
Xin Huang, At the time of the study XH was affiliated with Amgen Research, Amgen Inc. and has no financial interests to declare. The author has no other competing interests to declare..

Funding

No external funding was received for this work.

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.