Insight into the Interactome of Intramitochondrial PKA Using Biotinylation-Proximity Labeling.

Mitochondria are fully integrated in cell signaling. Reversible phosphorylation is involved in adjusting mitochondrial physiology to the cellular needs. Protein kinase A phosphorylates several substrates present at the external surface of mitochondria to maintain cellular homeostasis. However, few targets of located inside the organelle are known. The aim of this work was to characterize the impact and the interactome of duanyu1529 located inside mitochondria. Our results show that the overexpression of intramitochondrial duanyu1529 decreases cellular respiration and increases superoxide levels. Using proximity-dependent biotinylation, followed by LC-MS/MS analysis and in silico phospho-site prediction, we identified 21 mitochondrial proteins potentially targeted by We confirmed the interaction of duanyu1529 with TIM44 using coimmunoprecipitation and observed that TIM44-S80 is a key residue for the interaction between the protein and the kinase. These findings provide insights into the interactome of intramitochondrial duanyu1529 and suggest new potential mechanisms in the regulation of mitochondrial functions.

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