Investigating the molecular mechanisms of protein deCoAlation

辅酶A（Coenzyme A，CoA）是所有活细胞所必需的。1953年，F.李普曼因发现辅酶A及其在中间代谢中的重要性而获得诺贝尔奖。自这一里程碑式的发现以来，CoA及其衍生物（乙酰CoA，丙二酰CoA，HMG CoA等）的许多功能已被揭示，包括它们在脂肪酸合成和氧化，ATP生产，胆固醇和乙酰胆碱生物合成，氨基酸降解以及通过蛋白质乙酰化调节基因表达和细胞代谢中的重要性。辅酶A生物合成或辅酶A衍生物体内平衡失调与多种人类病理学有关，包括代谢紊乱、心脏肥大、癌症和神经退行性疾病。Gout教授的研究对哺乳动物辅酶A合酶的分子克隆和特征鉴定至关重要，并确定了这种酶的突变与帕金森氏症样神经退行性疾病（NBIA，神经退行性疾病伴脑铁积累）的侵袭性形式有关。最近，Gout教授开创了一个新的研究领域，即蛋白质结合和CoA的抗氧化功能。他发起并领导了一个关于这一新兴和重要研究领域的高知名度国际联盟。这些合作努力对于证明蛋白质CoAlation是由氧化剂和代谢应激在细菌和哺乳动物细胞中诱导的可逆翻译后修饰是必不可少的。在已建立的联盟的框架内，已开发出独特的试剂和方法，并证明对于鉴定细胞和组织中CoA修饰的蛋白质以及揭示蛋白质CoAlation的广泛性质至关重要。蛋白质结合被证明可以调节修饰蛋白质的亚细胞定位、酶活性和功能。在真核和原核细胞中广泛的蛋白质CoAlation的鉴定，以及这种翻译后修饰的可逆性质表明，还应该存在起介导CoA从修饰的蛋白质中去除的作用的酶。我们将这些酶称为CoAredoxins。这项研究提案利用了新的发现和新开发的方法，并将进一步推进蛋白质CoAlation的研究领域，并定义CoA作为细胞对氧化和代谢应激反应的重要抗氧化剂的作用。主要研究问题的目的是：（i）从细菌和哺乳动物细胞中鉴定和功能表征CoAredoxins;（ii）开发新的研究工具和方法，用于研究氧化还原调节和信号传导中的蛋白质CoAlation。这些问题的解决将揭示蛋白质去聚的分子基础，从而为CoA在细胞代谢和氧化还原调节中的基础生物学提供信息。这项工作将为阐明蛋白质结合在健康和疾病中的作用奠定基础。

Coenzyme A (CoA) is essential for all living cells. In 1953, F. Lipmann received a Nobel prize for the discovery of CoA and its importance in intermediary metabolism. Since this landmark discovery, numerous functions of CoA and its derivatives (Acetyl CoA, Malonyl CoA, HMG CoA among others) have been revealed, including their significance in the synthesis and oxidation of fatty acids, ATP production, biosynthesis of cholesterol and acetylcholine, degradation of amino acids and the regulation of gene expression and cellular metabolism via protein acetylation. Dysregulation of CoA biosynthesis or CoA derivatives homoeostasis is associated with various human pathologies, including metabolic disorders, cardiac hypertrophy, cancer and neurodegeneration.Prof Gout's research was central to molecular cloning and characterisation of mammalian CoA synthase, and the identification of mutations in this enzyme associated with an aggressive form of a Parkinson's-like neurodegeneration (NBIA, neurodegeneration with brain iron accumulation). Recently, Prof Gout pioneered a new field of research on protein CoAlation and antioxidant function of CoA. He has initiated and led a high profile international consortium on this emerging and significant area of research. These collaborative efforts were essential for demonstrating that protein CoAlation is a reversible post-translational modification induced in bacteria and mammalian cells by oxidising agents and metabolic stress. In the frame of the established consortium, unique reagents and methodologies have been developed and proved to be critical for identifying CoA-modified proteins in cells and tissues, and revealing a widespread nature of protein CoAlation. Protein CoAlation was shown to regulate the subcellular localisation, enzymatic activity and function of modified proteins. The identification of extensive protein CoAlation in eukaryotic and prokaryotic cells, and the reversible nature of this post-translational modification suggest there should also be enzymes which function to mediate the removal of CoA from modified proteins. We have termed these enzymes CoAredoxins. This research proposal leverages novel findings and newly developed methodologies, and will further advance the field of research on protein CoAlation and define the role of CoA as an important antioxidant in cellular response to oxidative and metabolic stress. Key research questions are aimed at: i) the identification and functional characterisation of CoAredoxins from bacteria and mammalian cells; and (ii) the development of novel research tools and methodologies for studying protein CoAlation in redox regulation and signalling. Answering these questions will reveal the molecular basis of protein deCoAlation and thus inform on the fundamental biology of CoA in cellular metabolism and redox regulation. This work will lay the foundation for delineating the role of protein CoAlation in health and disease.