Structural basis of TRPC4 regulation by calmodulin and pharmacological agents

Abstract
Data availability
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Abstract

Canonical transient receptor potential channels (TRPC) are involved in receptor-operated and/or store-operated Ca²⁺signaling. Inhibition of TRPCs by small molecules was shown to be promising in treating renal diseases. In cells, the channels are regulated by calmodulin. Molecular details of both calmodulin and drug binding have remained elusive so far. Here we report structures of TRPC4 in complex with three pyridazinone-based inhibitors and calmodulin. The structures reveal that all the inhibitors bind to the same cavity of the voltage-sensing-like domain and allow us to describe how structural changes from the ligand binding site can be transmitted to the central ion-conducting pore of TRPC4. Calmodulin binds to the rib helix of TRPC4, which results in the ordering of a previously disordered region, fixing the channel in its closed conformation. This represents a novel calmodulin-induced regulatory mechanism of canonical TRP channels.

Data availability

The atomic coordinates and cryo-EM maps for TRPC4DR in complex with inhibitors, calmodulin and for TRPC4DR in LMNG are available at the Protein Data Bank (PDB) and Electron Microscopy Data Bank (EMDB) databases, under the accession numbers PBD 7B0S and EMD-11970 (TRPC4-GFB8438), PBD 7B16 and EMD-11979 (TRPC4-GFB9289); PBD 7B05 and EMD-11957 (TRPC4-GFB8749); PBD 7B1G and EMD-11985 (TRPC4-Calmodulin) and PBD 7B0J and EMD-11968 (TRPC4-apo in LMNG).

The following data sets were generated

1. Raunser S et al
(2020) TRPC4-GFB8438
PDB, 7B0S.

https://www.rcsb.org/structure/7B0S
1. Raunser S et al
(2020) TRPC4-GFB9289
PDB, 7B16.

https://www.rcsb.org/structure/7B16
1. Raunser S et al
(2020) TRPC4-GFB8749
PDB, 7B05.

https://www.rcsb.org/structure/7B05
1. Raunser S et al
(2020) TRPC4-Calmodulin
PDB, 7B1G.

https://www.rcsb.org/structure/7B1G
1. Raunser S et al
(2020) TRPC4-apo in LMNG
PDB, 7B0J.

https://www.rcsb.org/structure/7B0J
1. Raunser S et al
(2020) TRPC4-GFB8438
EMDB, 11970.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-11970
1. Raunser S et al
(2020) TRPC4-GFB9289
EMDB, 11979.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-11979
1. Raunser S et al
(2020) TRPC4-GFB8749
EMDB, 11957.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-11957
1. Raunser S et al
(2020) TRPC4-Calmodulin
EMDB, 11985.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-11985
1. Raunser S et al
(2020) TRPC4-Calmodulin
EMDB, 11968.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-11968

Article and author information

Author details

Deivanayagabarathy Vinayagam

Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology, Dortmund, Germany

Competing interests
No competing interests declared.
Dennis Quentin

Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology, Dortmund, Germany

Competing interests
No competing interests declared.
Jing Yu-Strzelczyk

Department of Neurophysiology, Julius-Maximilians-Universität Würzburg, Würzburg, Germany

Competing interests
No competing interests declared.
Oleg Sitsel

Department of Structural Biochemistry, Max Planck Institute for Molecular Physiology, Dortmund, Germany

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0002-4496-7489
Felipe Merino

Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology, Dortmund, Germany

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0003-4166-8747
Markus Stabrin

Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology, Dortmund, Germany

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0003-0191-6419
Oliver Hofnagel

Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology, Dortmund, Germany

Competing interests
No competing interests declared.
Maolin Yu

Research Department, Goldfinch Bio Inc, Boston, United States

Competing interests
Maolin Yu, The author is or was a shareholder of Goldfinch Bio..
Mark W Ledeboer

Research Department, Goldfinch Bio Inc, Boston, United States

Competing interests
Mark W Ledeboer, The author is or was a shareholder of Goldfinch Bio..
Georg Nagel

Department of Biology, Institute for Molecular Plant Physiology and Biophysics, Biocenter, Julius-Maximilians-Universität Würzburg, Würzburg, Germany

Competing interests
No competing interests declared.
Goran Malojcic

Research Department, Goldfinch Bio Inc, Boston, United States

Competing interests
Goran Malojcic, The author is or was a shareholder of Goldfinch Bio..
Stefan Raunser

Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology, Dortmund, Germany

For correspondence
stefan.raunser@mpi-dortmund.mpg.de

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-9373-3016

Funding

Max-Planck-Gesellschaft

Stefan Raunser

Deutsche Forschungsgemeinschaft (TR240)

Georg Nagel

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.