Introduction and Aim Mitochondrial cristae are dynamic compartments with distinct structures that contribute to respiratory efficiency. Optic Atrophy 1 (Opa1), a dynamin-related inner mitochondrial membrane (IMM) protein, is a central regulator of cristae shape that is retrieved in oligomeric complexes modulated by metabolic changes, but whether and how Opa1 connects fuel availability to cristae dynamics is unexplored. Methods and Results To address this question, we generated a catalogue of the dynamic Opa1 interactome upon changes in fuel availability. We engineered an active, correctly localized and oligomerized Opa1-TurboID (Opa1TID) chimeric protein that complements mitochondrial ultrastructure and fusion of Opa1-/- cells. Using Opa1TID, we identified by liquid chromatography-label free proteomics the Opa1 interactome in Murine Embryonic Fibroblasts exposed to glucose, starvation, fatty acids, amino acids, or a complete cell culture medium mimicking human plasma. Among the identified 231 bona fide mitochondrial proteins of Opa1TID interactors, 44 unique interactors were significantly enriched in distinct metabolic conditions, pinpointing to fatty acid, amino acid and solute carriers as potential relays between changing fuels and cristae structure. Furthermore, we engineered a conditional flox-stop Opa1TID murine model that we crossed to the tamoxifen inducible, cardiac specific alpha-MHC-MerCreMer mouse to develop a heart specific, inducible Opa1TID mouse model (Opa1iHTID) to study the Opa1 interactome in cardiac pathophysiology. Conclusion This discovery of the carbon source-dependent Opa1TID interactome indicates an axis between fuel availability and Opa1-mediated cristae dynamics and pinpoints metabolic enzymes that can relay individual fuel source to cristae biogenesis machinery. The Opa1iHTID mouse will shed light on cardiac Opa1 interactors in vivo, offering a metabolic strategy to prevent pathogenic Opa1-mediated cristae remodeling during heart IR injury or failure.
The metabolically dependent interactomes of the mitochondrial cristae-shaping protein Opa1 in cardiac pathophysiology
Jonathan LambertInvestigation
;Luca ScorranoSupervision
In corso di stampa
Abstract
Introduction and Aim Mitochondrial cristae are dynamic compartments with distinct structures that contribute to respiratory efficiency. Optic Atrophy 1 (Opa1), a dynamin-related inner mitochondrial membrane (IMM) protein, is a central regulator of cristae shape that is retrieved in oligomeric complexes modulated by metabolic changes, but whether and how Opa1 connects fuel availability to cristae dynamics is unexplored. Methods and Results To address this question, we generated a catalogue of the dynamic Opa1 interactome upon changes in fuel availability. We engineered an active, correctly localized and oligomerized Opa1-TurboID (Opa1TID) chimeric protein that complements mitochondrial ultrastructure and fusion of Opa1-/- cells. Using Opa1TID, we identified by liquid chromatography-label free proteomics the Opa1 interactome in Murine Embryonic Fibroblasts exposed to glucose, starvation, fatty acids, amino acids, or a complete cell culture medium mimicking human plasma. Among the identified 231 bona fide mitochondrial proteins of Opa1TID interactors, 44 unique interactors were significantly enriched in distinct metabolic conditions, pinpointing to fatty acid, amino acid and solute carriers as potential relays between changing fuels and cristae structure. Furthermore, we engineered a conditional flox-stop Opa1TID murine model that we crossed to the tamoxifen inducible, cardiac specific alpha-MHC-MerCreMer mouse to develop a heart specific, inducible Opa1TID mouse model (Opa1iHTID) to study the Opa1 interactome in cardiac pathophysiology. Conclusion This discovery of the carbon source-dependent Opa1TID interactome indicates an axis between fuel availability and Opa1-mediated cristae dynamics and pinpoints metabolic enzymes that can relay individual fuel source to cristae biogenesis machinery. The Opa1iHTID mouse will shed light on cardiac Opa1 interactors in vivo, offering a metabolic strategy to prevent pathogenic Opa1-mediated cristae remodeling during heart IR injury or failure.
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