puc-header

Gating Movements and Ion Permeation in HCN4 Pacemaker Channels

68 Pages Posted: 22 Dec 2020 Publication Status: Published

See all articles by Andrea Saponaro

Andrea Saponaro

University of Milan - Department of Biosciences

Daniel Bauer

TU-Darmstadt - Department of Biology

M. Hunter Giese

Columbia University - Department of Physiology and Cellular Biophysics

Paolo Swuec

University of Milan - Department of Biosciences

Alessandro Porro

University of Milan - Department of Biosciences

Federica Gasparri

University of Milan - Department of Biosciences

Atiyeh Sadat Sharifzadeh

University of Milan - Department of Biosciences

Antonio Chaves-Sanjuan

University of Milan - Department of Biosciences

Laura Alberio

Newcastle University - Biosciences Institute

Giacomo Parisi

Instituto Italiano di Tecnologia - Center for Life Nano Science

Gabriele Cerutti

Columbia University - Department of Biochemistry and Molecular Biophysics

Oliver Clarke

Columbia University - Department of Physiology and Cellular Biophysics

Kay Hamacher

TU-Darmstadt - Department of Biology

Henry M. Colecraft

Columbia University - Department of Physiology and Cellular Biophysics

Filippo Mancia

Columbia University - Department of Physiology

Wayne A. Hendrickson

Columbia University - Department of Physiology and Cellular Biophysics

Steven A. Siegelbaum

Columbia University - Department of Neuroscience

Dario DiFrancesco

University of Milan - Department of Biosciences

Martino Bolognesi

University of Milan - Department of Biosciences

Gerhard Thiel

TU-Darmstadt - Department of Biology

Bina Santoro

Columbia University - Department of Neuroscience

Anna Moroni

University of Milan - Department of Biosciences

More...

Abstract

HCN channels underlie hyperpolarization-activated cation current If/Ih that controls automaticity in cardiac and neuronal pacemaker cells. We present cryo-EM structures of HCN4 in the presence/absence of bound cAMP, displaying the pore domain in closed and open conformations. Analysis of cAMP-bound and -unbound structures sheds light on how ligand-induced transitions in the channel cytosolic portion mediate the effect of cAMP on channel gating, and highlights the regulatory role of a Mg2+ coordination site formed between the C-linker and the S4-S5 linker. Comparison of open/closed pore states shows that the cytosolic gate opens through concerted movements of the S5 and S6 transmembrane helices. In combination with molecular dynamics, the open pore structures provide insights into the mechanisms of K+/Na+ permeation, revealing distinctive ion-binding dependent adaptation in the selectivity filter. Our results contribute new mechanistic understanding on HCN channel gating, cyclic nucleotide-dependent modulation, and ion permeation.

Keywords: HCN4, pore, gating, selectivity, permeation, cAMP, cryo-EM, molecular dynamics, pacemaker

Suggested Citation

Saponaro, Andrea and Bauer, Daniel and Giese, M. Hunter and Swuec, Paolo and Porro, Alessandro and Gasparri, Federica and Sharifzadeh, Atiyeh Sadat and Chaves-Sanjuan, Antonio and Alberio, Laura and Parisi, Giacomo and Cerutti, Gabriele and Clarke, Oliver and Hamacher, Kay and Colecraft, Henry M. and Mancia, Filippo and Hendrickson, Wayne A. and Siegelbaum, Steven A. and DiFrancesco, Dario and Bolognesi, Martino and Thiel, Gerhard and Santoro, Bina and Moroni, Anna, Gating Movements and Ion Permeation in HCN4 Pacemaker Channels. Available at SSRN: https://ssrn.com/abstract=3753801 or http://dx.doi.org/10.2139/ssrn.3753801
This version of the paper has not been formally peer reviewed.

Andrea Saponaro

University of Milan - Department of Biosciences

Via Festa del Perdono, 7
Milan, 20122
Italy

Daniel Bauer

TU-Darmstadt - Department of Biology

Universitaets- und Landesbibliothek Darmstadt
Magdalenenstrasse 8
Darmstadt, Hesse D-64289
Germany

M. Hunter Giese

Columbia University - Department of Physiology and Cellular Biophysics ( email )

3022 Broadway
New York, NY 10027
United States

Paolo Swuec

University of Milan - Department of Biosciences

via Festa del Perdono, 7
Milano
Italy

Alessandro Porro

University of Milan - Department of Biosciences ( email )

Italy

Federica Gasparri

University of Milan - Department of Biosciences ( email )

Italy

Atiyeh Sadat Sharifzadeh

University of Milan - Department of Biosciences ( email )

Italy

Antonio Chaves-Sanjuan

University of Milan - Department of Biosciences

via Festa del Perdono, 7
Milano
Italy

Laura Alberio

Newcastle University - Biosciences Institute ( email )

Newcastle upon Tyne
United Kingdom

Giacomo Parisi

Instituto Italiano di Tecnologia - Center for Life Nano Science ( email )

Gabriele Cerutti

Columbia University - Department of Biochemistry and Molecular Biophysics ( email )

3022 Broadway
New York, NY 10027
United States

Oliver Clarke

Columbia University - Department of Physiology and Cellular Biophysics ( email )

3022 Broadway
New York, NY 10027
United States

Kay Hamacher

TU-Darmstadt - Department of Biology ( email )

Universitaets- und Landesbibliothek Darmstadt
Magdalenenstrasse 8
Darmstadt, Hesse D-64289
Germany

Henry M. Colecraft

Columbia University - Department of Physiology and Cellular Biophysics ( email )

3022 Broadway
New York, NY 10027
United States

Filippo Mancia

Columbia University - Department of Physiology ( email )

1150 St. Nicholas Ave., Room 508
Russ Berrie Pavilion
New York, NY 10032
United States

Wayne A. Hendrickson

Columbia University - Department of Physiology and Cellular Biophysics ( email )

3022 Broadway
New York, NY 10027
United States

Steven A. Siegelbaum

Columbia University - Department of Neuroscience ( email )

615 West 131st Street, 6th floor
Studebaker Building, MC 8765
New York, NY 10027
United States

Dario DiFrancesco

University of Milan - Department of Biosciences ( email )

Italy

Martino Bolognesi

University of Milan - Department of Biosciences

via Festa del Perdono, 7
Milano
Italy

Gerhard Thiel

TU-Darmstadt - Department of Biology ( email )

Universitaets- und Landesbibliothek Darmstadt
Magdalenenstrasse 8
Darmstadt, Hesse D-64289
Germany

Bina Santoro

Columbia University - Department of Neuroscience ( email )

615 West 131st Street, 6th floor
Studebaker Building, MC 8765
New York, NY 10027
United States

Anna Moroni (Contact Author)

University of Milan - Department of Biosciences ( email )

Italy

Click here to go to Cell.com

Paper statistics

Downloads
35
Abstract Views
607
PlumX Metrics