Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK.
Biological Mass Spectrometry & Proteomics Cell Biology, MRC Laboratory of Molecular Biology, Cambridge, CB2 0QH, UK.
Mol Cell Biochem. 2019 Nov;461(1-2):91-102. doi: 10.1007/s11010-019-03593-w. Epub 2019 Aug 2.
Peroxiredoxins (Prdxs) are antioxidant enzymes that catalyse the breakdown of peroxides and regulate redox activity in the cell. Peroxiredoxin 5 (Prdx5) is a unique member of Prdxs, which displays a wider subcellular distribution and substrate specificity and exhibits a different catalytic mechanism when compared to other members of the family. Here, the role of a key metabolic integrator coenzyme A (CoA) in modulating the activity of Prdx5 was investigated. We report for the first time a novel mode of Prdx5 regulation mediated via covalent and reversible attachment of CoA (CoAlation) in cellular response to oxidative and metabolic stress. The site of CoAlation in endogenous Prdx5 was mapped by mass spectrometry to peroxidatic cysteine 48. By employing an in vitro CoAlation assay, we showed that Prdx5 peroxidase activity is inhibited by covalent interaction with CoA in a dithiothreitol-sensitive manner. Collectively, these results reveal that human Prdx5 is a substrate for CoAlation in vitro and in vivo, and provide new insight into metabolic control of redox status in mammalian cells.
过氧化物酶（Prdxs）是一种抗氧化酶，可催化过氧化物的分解，并调节细胞内的氧化还原活性。过氧化物酶 5（Prdx5）是 Prdxs 的一个独特成员，与家族中的其他成员相比，它具有更广泛的亚细胞分布和底物特异性，并表现出不同的催化机制。在这里，研究了关键代谢整合辅酶 A（CoA）在调节 Prdx5 活性中的作用。我们首次报道了一种新的 Prdx5 调节模式，该模式通过共价和可逆的 CoA 附着（CoAlation）来介导，以响应氧化和代谢应激。通过质谱法将 CoAlation 在内源性 Prdx5 中的位点映射到过氧物酶活性中心的半胱氨酸 48。通过体外 CoAlation 测定，我们表明 Prdx5 过氧化物酶活性可通过与 CoA 的共价相互作用以二硫苏糖醇敏感的方式被抑制。总之，这些结果表明人 Prdx5 是体外和体内 CoAlation 的底物，并为哺乳动物细胞中氧化还原状态的代谢控制提供了新的见解。
