Phosphoproteomics reveals conserved exercise‐stimulated signaling and AMPK regulation of store‐operated calcium entry

• Published in: The EMBO journal Vol. 38; no. 24; pp. e102578 - n/a
• Main Authors: Nelson, Marin E, Parker, Benjamin L, Burchfield, James G, Hoffman, Nolan J, Needham, Elise J, Cooke, Kristen C, Naim, Timur, Sylow, Lykke, Ling, Naomi XY, Francis, Deanne, Norris, Dougall M, Chaudhuri, Rima, Oakhill, Jonathan S, Richter, Erik A, Lynch, Gordon S, Stöckli, Jacqueline, James, David E
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 16.12.2019; Blackwell Publishing Ltd; John Wiley and Sons Inc
• Subjects: Adaptation; AMP-Activated Protein Kinases - metabolism; AMPK; Animal models; Animals; Calcium; Calcium (intracellular); Calcium - metabolism; Calcium Channels - metabolism; Calcium Signaling - physiology; Crosstalk; Drosophila; EMBO21; EMBO31; EMBO37; exercise; Female; Humans; Insects; Intracellular; Intracellular signalling; Kinases; Male; Mass spectrometry; Mass spectroscopy; Membrane Proteins; Mice; Muscle contraction; Muscle, Skeletal - metabolism; Muscles; Muscular function; Musculoskeletal system; Phosphorylation; Physical training; Physiology; Protein Conformation; Proteins; Proteomics - methods; Rats; Rats, Wistar; Regulators; Resource; Signal Transduction; Signaling; Skeletal muscle; Species; STIM1; STIM1 protein; Stromal Interaction Molecule 1 - chemistry; Stromal Interaction Molecule 1 - genetics; Stromal Interaction Molecule 1 - metabolism; Substrates; Treadmills; AMPK; exercise; calcium; STIM1; phosphorylation
• ISSN: 0261-4189; 1460-2075
• DOI: 10.15252/embj.2019102578

Exercise stimulates cellular and physiological adaptations that are associated with widespread health benefits. To uncover conserved protein phosphorylation events underlying this adaptive response, we performed mass spectrometry‐based phosphoproteomic analyses of skeletal muscle from two widely used rodent models: treadmill running in mice and in situ muscle contraction in rats. We overlaid these phosphoproteomic signatures with cycling in humans to identify common cross‐species phosphosite responses, as well as unique model‐specific regulation. We identified > 22,000 phosphosites, revealing orthologous protein phosphorylation and overlapping signaling pathways regulated by exercise. This included two conserved phosphosites on stromal interaction molecule 1 (STIM1), which we validate as AMPK substrates. Furthermore, we demonstrate that AMPK‐mediated phosphorylation of STIM1 negatively regulates store‐operated calcium entry, and this is beneficial for exercise in Drosophila . This integrated cross‐species resource of exercise‐regulated signaling in human, mouse, and rat skeletal muscle has uncovered conserved networks and unraveled crosstalk between AMPK and intracellular calcium flux. Synopsis Exercise stimulates cellular and physiological adaptations associated with widespread health benefits, however the underlying signalling events remain unclear. Here, integrated cross‐species analysis of exercise‐regulated protein phosphorylation in the skeletal muscle uncovers conserved signaling networks including crosstalk between AMPK and intracellular calcium flux. Phospoproteomic analysis of human, rat and mouse skeletal muscle identifies > 5,000 exercise‐regulated phosphosites. Cross‐species integration reveals conserved pathways and a compendium of potential exercise regulators. AMPK phosphorylates stromal interaction molecule 1 (STIM1) and inhibits skeletal muscle store‐operated calcium entry. Inhibition of STIM1 improves exercise tolerance and delays fatigue in Drosophila . Graphical Abstract Global analysis of contraction‐regulated protein phosphorylation in skeletal muscle uncovers signalling nodes conserved between mouse, rat and human.