Mechanism of adrenergic CaV1.2 stimulation revealed by proximity proteomics
Open accessAbstract
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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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.
- ORCID
- Venue
- Nature
- Published
- 2020-01-22
- Volume
- 577
- Issue
- 7792
- Pages
- 695-700
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC7018383
- 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
- green
- www.ncbi.nlm.nih.gov
- 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
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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