Investigating the molecular mechanisms of protein deCoAlation

辅酶A(CoA)是所有活细胞所必需的。1953年，F·利普曼因发现辅酶A及其在中间代谢中的重要性而获得诺贝尔奖。自从这一里程碑式的发现以来，辅酶A及其衍生物(乙酰辅酶A、丙二酰辅酶A、HMG辅酶A等)的许多功能已经被揭示，包括它们在脂肪酸的合成和氧化、ATP的产生、胆固醇和乙酰胆碱的生物合成、氨基酸的降解以及通过蛋白质乙酰化调节基因表达和细胞代谢方面的重要意义。辅酶A生物合成或辅酶A衍生物的平衡失调与各种人类病理有关，包括代谢紊乱、心脏肥大、癌症和神经退行性变。Gout教授的研究是哺乳动物辅酶A合成酶分子克隆和表征的核心，并确定这种酶的突变与一种侵袭性的帕金森样神经退行性变(NBIA，神经退行性变伴脑铁积聚)有关。最近，Gout教授开创了蛋白质结合和CoA抗氧化功能研究的新领域。他在这一新兴和重要的研究领域发起并领导了一个备受瞩目的国际财团。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至关重要。在已建立的联合体的框架内，已经开发出独特的试剂和方法，并被证明对于在细胞和组织中识别CoA修饰的蛋白质和揭示蛋白质CoAlation的广泛性质至关重要。蛋白质相互作用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发现了广泛的蛋白质CoA结合，以及这种翻译后修饰的可逆性，这表明也应该有酶来调节修饰蛋白质中CoA的去除。我们将这些酶命名为辅酶还蛋白。这项研究方案充分利用了新的发现和新开发的方法，将进一步推动蛋白质相互作用的研究领域，并确定辅酶A在细胞对氧化和代谢应激反应中作为重要抗氧化剂的作用。主要的研究问题是：i)细菌和哺乳动物细胞中CoAredoxins的鉴定和功能特征；以及(Ii)开发新的研究工具和方法来研究氧化还原调节和信号传递中的蛋白质相互作用。回答这些问题将揭示蛋白质解离的分子基础，从而了解辅酶A在细胞代谢和氧化还原调节中的基础生物学。这项工作将为阐明蛋白质相互作用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.