BPS2026 – Mitochondrial coenzyme A import and metabolism

The solute carrier SLC25 family is the largest group of mitochondrial metabolite transporters in eukaryotes that support metabolite translocation across the mitochondrial membrane. These metabolites include a wide range of nucleotides, amino acids, carboxylic acids, and cofactors. However, molecular mechanisms for metabolite translocation are unclear, and a large fraction of these SLC25 transporters is poorly characterized. Here, we will focus on mitochondrial coenzyme A (CoASH). CoASH is an essential cofactor that enriches in the mitochondrial matrix supporting mitochondrial bioenergetics. We developed an optimized LC-MS-based extraction and profiling workflow that enables robust detection of 33 cellular and 23 mitochondrial acyl-CoA species from cultured human cells. Using this approach, we identified SLC25A16 and SLC25A42 as key transporters mediating mitochondrial import of free CoASH, supporting TCA cycle and fatty acid oxidation. And de novo CoA synthesis occurs predominantly in the cytosol to support lipid anabolism. Together, these findings reveal that mitochondrial acyl-CoA compartmentalization orchestrates the spatial segregation of anabolic and catabolic metabolism, providing new insights into the pathophysiology of inborn errors of CoA metabolism. This abstract is based on data previously published in Liu et al. Nature Metabolism 2025.