Liu Ran, Zhang Zihan, Kyaw Aye K, Grabińska Kariona A, Shah Hardik, Shen Hongying
Cellular and Molecular Physiology Department, Yale School of Medicine, New Haven, CT, USA.
Systems Biology Institute, Yale West Campus, West Haven, CT, USA.
Nat Metab. 2025 Sep 9. doi: 10.1038/s42255-025-01358-y.
The essential cofactor coenzyme A (CoASH) and its thioester derivatives (acyl-CoAs) have pivotal roles in cellular metabolism. However, the mechanism by which different acyl-CoAs are accurately partitioned into different subcellular compartments to support site-specific reactions, and the physiological impact of such compartmentalization, remain poorly understood. Here, we report an optimized liquid chromatography-mass spectrometry-based pan-chain acyl-CoA extraction and profiling method that enables a robust detection of 33 cellular and 23 mitochondrial acyl-CoAs from cultured human cells. We reveal that SLC25A16 and SLC25A42 are critical for mitochondrial import of free CoASH. This CoASH import process supports an enriched mitochondrial CoA pool and CoA-dependent pathways in the matrix, including the high-flux TCA cycle and fatty acid oxidation. Despite a small fraction of the mitochondria-localized CoA synthase COASY, de novo CoA biosynthesis is primarily cytosolic and supports cytosolic lipid anabolism. This mitochondrial acyl-CoA compartmentalization enables a spatial regulation of anabolic and energy-related catabolic processes, which promises to shed light on pathophysiology in the inborn errors of CoA metabolism.
必需辅因子辅酶A（CoASH）及其硫酯衍生物（酰基辅酶A）在细胞代谢中起关键作用。然而，不同的酰基辅酶A如何准确地分配到不同的亚细胞区室以支持位点特异性反应的机制，以及这种区室化的生理影响，仍知之甚少。在这里，我们报告了一种基于液相色谱-质谱的优化的全链酰基辅酶A提取和分析方法，该方法能够从培养的人类细胞中可靠地检测33种细胞内和23种线粒体内的酰基辅酶A。我们发现SLC25A16和SLC25A42对游离CoASH的线粒体导入至关重要。这种CoASH导入过程支持线粒体基质中丰富的辅酶A池和依赖辅酶A的途径，包括高通量的三羧酸循环和脂肪酸氧化。尽管线粒体定位的辅酶A合酶COASY占比小，但从头合成辅酶A主要发生在胞质溶胶中，并支持胞质脂质合成代谢。这种线粒体酰基辅酶A区室化能够对合成代谢和能量相关的分解代谢过程进行空间调节，这有望为深入了解辅酶A代谢先天性缺陷中的病理生理学提供线索。
