Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.


Sci. Signal., 4 June 2013
Vol. 6, Issue 278, p. rs11
[DOI: 10.1126/scisignal.2003506]


In Vivo Phosphoproteomics Analysis Reveals the Cardiac Targets of β-Adrenergic Receptor Signaling

Alicia Lundby1,2,3*, Martin N. Andersen3, Annette B. Steffensen3, Heiko Horn1, Christian D. Kelstrup1, Chiara Francavilla1, Lars J. Jensen1, Nicole Schmitt3, Morten B. Thomsen3, and Jesper V. Olsen1*

1 Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3b, DK-2200 Copenhagen, Denmark.
2 The Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
3 The Danish National Research Foundation Centre for Cardiac Arrhythmia, DK-2200 Copenhagen N, Denmark.

Abstract: β-Blockers are widely used to prevent cardiac arrhythmias and to treat hypertension by inhibiting β-adrenergic receptors (βARs) and thus decreasing contractility and heart rate. βARs initiate phosphorylation-dependent signaling cascades, but only a small number of the target proteins are known. We used quantitative in vivo phosphoproteomics to identify 670 site-specific phosphorylation changes in murine hearts in response to acute treatment with specific βAR agonists. The residues adjacent to the regulated phosphorylation sites exhibited a sequence-specific preference (R-X-X-pS/T), and integrative analysis of sequence motifs and interaction networks suggested that the kinases AMPK (adenosine 5'-monophosphate–activated protein kinase), Akt, and mTOR (mammalian target of rapamycin) mediate βAR signaling, in addition to the well-established pathways mediated by PKA (cyclic adenosine monophosphate–dependent protein kinase) and CaMKII (calcium/calmodulin-dependent protein kinase type II). We found specific regulation of phosphorylation sites on six ion channels and transporters that mediate increased ion fluxes at higher heart rates, and we showed that phosphorylation of one of these, Ser92 of the potassium channel KV7.1, increased current amplitude. Our data set represents a quantitative analysis of phosphorylated proteins regulated in vivo upon stimulation of seven-transmembrane receptors, and our findings reveal previously unknown phosphorylation sites that regulate myocardial contractility, suggesting new potential targets for the treatment of heart disease and hypertension.

* Corresponding author. E-mail: alicia.lundby{at} (A.L.); jesper.olsen{at} (J.V.O.)

Citation: A. Lundby, M. N. Andersen, A. B. Steffensen, H. Horn, C. D. Kelstrup, C. Francavilla, L. J. Jensen, N. Schmitt, M. B. Thomsen, J. V. Olsen, In Vivo Phosphoproteomics Analysis Reveals the Cardiac Targets of β-Adrenergic Receptor Signaling. Sci. Signal. 6, rs11 (2013).

Read the Full Text

Status of Large-scale Analysis of Post-translational Modifications by Mass Spectrometry.
J. V. Olsen and M. Mann (2013)
Mol. Cell. Proteomics 12, 3444-3452
   Abstract »    Full Text »    PDF »
Development of heart failure is independent of K+ channel-interacting protein 2 expression.
T. Speerschneider, S. Grubb, A. Metoska, S.-P. Olesen, K. Calloe, and M. B. Thomsen (2013)
J. Physiol. 591, 5923-5937
   Abstract »    Full Text »    PDF »
Phosphorylation sites required for regulation of cardiac calcium channels in the fight-or-flight response.
Y. Fu, R. E. Westenbroek, T. Scheuer, and W. A. Catterall (2013)
PNAS 110, 19621-19626
   Abstract »    Full Text »    PDF »
Phosphoproteomics Study Based on In Vivo Inhibition Reveals Sites of Calmodulin-Dependent Protein Kinase II Regulation in the Heart.
A. Scholten, C. Preisinger, E. Corradini, V. J. Bourgonje, M. L. Hennrich, T. A. B. van Veen, P. D. Swaminathan, M.-L. Joiner, M. A. Vos, M. E. Anderson, et al. (2013)
JAHA 2, e000318
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882