关键词： BioID2 TIM44 mitochondria protein kinase A proteomics serine/threonine phosphoprediction

Mesh ： Biotinylation / methods Cell Respiration / physiology Cross-Linking Reagents / chemistry Cyclic AMP-Dependent Protein Kinases / chemistry metabolism HEK293 Cells HeLa Cells Humans Immunoprecipitation / methods Microscopy, Fluorescence Mitochondria / chemistry metabolism Mitochondrial Proteins / chemistry metabolism Protein Binding Protein Interaction Mapping / methods Protein Processing, Post-Translational Signal Transduction Staining and Labeling / methods

来  源：   DOI：10.3390/ijms21218283   PDF(Sci-hub)   PDF(Pubmed)

Abstract：
Mitochondria are fully integrated in cell signaling. Reversible phosphorylation is involved in adjusting mitochondrial physiology to the cellular needs. Protein kinase A (PKA) phosphorylates several substrates present at the external surface of mitochondria to maintain cellular homeostasis. However, few targets of PKA located inside the organelle are known. The aim of this work was to characterize the impact and the interactome of PKA located inside mitochondria. Our results show that the overexpression of intramitochondrial PKA 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 PKA. We confirmed the interaction of PKA 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 PKA and suggest new potential mechanisms in the regulation of mitochondrial functions.
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