Cited by 243Ion channel regulation and functionCardiac electrophysiology and arrhythmiasReceptor Mechanisms and Signaling

Mechanism of adrenergic CaV1.2 stimulation revealed by proximity proteomics

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Liu, Guoxia, Papa, Arianne, Katchman, Alexander N., Zakharov, Sergey I., Roybal, Daniel, Hennessey, Jessica A., Kushner, Jared, Yang, Lin +9 more·2020·Nature

Abstract

Increased cardiac contractility during the fight-or-flight response is caused by β-adrenergic augmentation of Ca V 1.2 voltage-gated calcium channels 1-4. However, this augmentation persists in transgenic murine hearts expressing mutant Ca V 1.2 α 1C and β subunits that can no longer be phosphorylated by protein kinase A-an essential downstream mediator of β-adrenergic signalling-suggesting that non-channel factors are also required. Here we identify the mechanism by which β-adrenergic agonists stimulate voltage-gated calcium channels. We express α 1C or β 2B subunits conjugated to ascorbate peroxidase 5 in mouse hearts, and use multiplexed quantitative proteomics 6,7 to track hundreds of proteins in the proximity of Ca V 1.2. We observe that the calcium-channel inhibitor Rad 8,9, a monomeric G protein, is enriched in the Ca V 1.2 microenvironment but is depleted during β-adrenergic stimulation. Phosphorylation by protein kinase A of specific serine residues on Rad decreases its affinity for β subunits and relieves constitutive inhibition of Ca V 1.2, observed as an increase in channel open probability. Expression of Rad or its homologue Rem in HEK293T cells also imparts stimulation of Ca V 1.3 and Ca V 2.2 by protein kinase A, revealing an evolutionarily conserved mechanism that confers adrenergic modulation upon voltage-gated calcium channels.

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Crossref · 10.1038/s41586-020-1947-z

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  • Proximity labelling assessed protein-protein interactions involved in β-adrenergic signalling of contractility in cardiac fight-or-flight responses and evaluation of regenerative stem cell therapy efficacy in post-infarct hearts.

    “proximity labelling to assess protein–protein interactions involved in β-adrenergic signalling of contractility in cardiac fight-or-flight responses124 and evaluation of regenerative stem cell therapy efficacy in post-infarct hearts125.”

    From A high-stringency blueprint of the human proteome (2020) · cites this paper as [124]

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Authors
Liu, G., Papa, A., Katchman, A. N., Zakharov, S. I., Roybal, D., Hennessey, J. A., Kushner, J., Yang, L., Chen, B.-X., Kushnir, A., Dangas, K., Gygi, S. P., Pitt, G. S., Colecraft, H. M., Ben-Johny, M., Kalocsay, M., & Marx, S. O.
Affiliations
  • Columbia University (US)
  • Harvard University (US)
  • Weill Cornell Medicine (US)
  • Columbia University Irving Medical Center (US)
Corresponding author
  • Kalocsay, Marian
  • Marx, Steven O.
Venue
Nature
Published
2020-01-22
Volume
577
Issue
7792
Pages
695-700
ISSN
  • 0028-0836 (ISSN-L)
  • 1476-4687
OpenAlex
W3003002159
Cited by
243
References
74
Citations by year
2026: 23 · 2025: 31 · 2024: 36 · 2023: 36 · 2022: 39 · 2021: 50 · 2020: 28
FWCI
12.41
Type
journal article
Language
en
Open access
Locations
  • Nature (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • Digital Access to Scholarship at Harvard (DASH) (Harvard University) (acceptedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
Funders
  • National Institutes of Health (UL1TR001873)
  • National Heart, Lung, and Blood Institute (R01HL113136, F31HL142178, T32HL120826, R01HL126735, R01HL121253, R01HL140934, R01HL146149, T32HL160520, T32HL007343, T32HL007854)
  • National Cancer Institute (P30CA013696)
  • National Center for Advancing Translational Sciences
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion channel regulation and function
  • Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Cardiac electrophysiology and arrhythmias
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
Keywords
Cav1.2, voltage-gated calcium channels, beta-adrenergic signaling, protein kinase A, REM, G proteins, channel phosphorylation, channel inhibition, cardiac contractility, proximity proteomics, ascorbate peroxidase, Cav1.3, CaV2.2, calcium current
MeSH terms
Protein Domains, Cyclic AMP, Animals, Female, Humans, Male, Myocardium, Phosphorylation, Receptors, Adrenergic, beta, Signal Transduction, Cyclic AMP-Dependent Protein Kinases, ras Proteins, Monomeric GTP-Binding Proteins, Calcium Channels, L-Type, Calcium Channels, N-Type, Heterotrimeric GTP-Binding Proteins, Protein Subunits, Proteomics, Mice, HEK293 Cells, Cellular Microenvironment

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