Global phosphoproteomic profiling reveals perturbed signaling in a mouse model of dilated cardiomyopathy


Kuzmanov, U, Guo, H, Buchsbaum, D, Cosme, J, Abbasi, C, Isserlin, R, Sharma, P ORCID: 0000-0002-5534-2417, Gramolini, A and Emili, A (2016) Global phosphoproteomic profiling reveals perturbed signaling in a mouse model of dilated cardiomyopathy. Proceedings of the National Academy of Sciences, 113 (44). pp. 12592-12597.

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Abstract

Phospholamban (PLN) plays a central role in Ca2+ homeostasis in cardiac myocytes through regulation of the sarco(endo)plasmic reticulum Ca2+-ATPase 2A (SERCA2A) Ca2+ pump. An inherited mutation converting arginine residue 9 in PLN to cysteine (R9C) results in dilated cardiomyopathy (DCM) in humans and transgenic mice, but the downstream signaling defects leading to decompensation and heart failure are poorly understood. Here we used precision mass spectrometry to study the global phosphorylation dynamics of 1,887 cardiac phosphoproteins in early affected heart tissue in a transgenic R9C mouse model of DCM compared with wild-type littermates. Dysregulated phosphorylation sites were quantified after affinity capture and identification of 3,908 phosphopeptides from fractionated whole-heart homogenates. Global statistical enrichment analysis of the differential phosphoprotein patterns revealed selective perturbation of signaling pathways regulating cardiovascular activity in early stages of DCM. Strikingly, dysregulated signaling through the Notch-1 receptor, recently linked to cardiomyogenesis and embryonic cardiac stem cell development and differentiation but never directly implicated in DCM before, was a prominently perturbed pathway. We verified alterations in Notch-1 downstream components in early symptomatic R9C transgenic mouse cardiomyocytes compared with wild type by immunoblot analysis and confocal immunofluorescence microscopy. These data reveal unexpected connections between stress-regulated cell signaling networks, specific protein kinases, and downstream effectors essential for proper cardiac function.

Item Type: Article
Uncontrolled Keywords: phospholamban, proteomic, bioinformatics, heart disease, signaling
Depositing User: Symplectic Admin
Date Deposited: 17 Oct 2016 09:39
Last Modified: 19 Jan 2023 07:29
DOI: 10.1073/pnas.1606444113
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3003794

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