Revisiting insulin-stimulated hydrogen peroxide dynamics reveals a cytosolic reductive shift in skeletal muscle

The intracellular redox state is crucial for insulin responses in peripheral tissues. Despite the longstanding belief that insulin signaling increases hydrogen peroxide (H2O2) production leading to reversible oxidation of cysteine thiols, evidence is inconsistent and rarely involves human tissues. I...

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Main Authors: Carlos Henríquez-Olguín, Samantha Gallero, Anita Reddy, Kaspar W. Persson, Farina L. Schlabs, Christian T. Voldstedlund, Gintare Valentinaviciute, Roberto Meneses-Valdés, Casper M. Sigvardsen, Bente Kiens, Edward T. Chouchani, Erik A. Richter, Thomas E. Jensen
Format: Article
Language:English
Published: Elsevier 2025-05-01
Series:Redox Biology
Online Access:http://www.sciencedirect.com/science/article/pii/S221323172500120X
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Summary:The intracellular redox state is crucial for insulin responses in peripheral tissues. Despite the longstanding belief that insulin signaling increases hydrogen peroxide (H2O2) production leading to reversible oxidation of cysteine thiols, evidence is inconsistent and rarely involves human tissues. In this study, we systematically investigated insulin-dependent changes in subcellular H2O2 levels and reversible cysteine modifications across mouse and human skeletal muscle models. Utilizing advanced redox tools—including genetically encoded H2O2 sensors and non-reducing immunoblotting—we consistently observed no increase in subcellular H2O2 levels following insulin stimulation. Instead, stoichiometric cysteine proteome analyses revealed a selective pro-reductive shift in cysteine modifications affecting insulin transduction related proteins, including Cys179 on GSK3β and Cys416 on Ras and Rab Interactor 2 (RIN2). Our findings challenge the prevailing notion that insulin promotes H2O2 generation in skeletal muscle and suggest that an insulin-stimulated pro-reductive shift modulates certain aspects of insulin signal transduction.
ISSN:2213-2317