Understand Glycation: The Sweet Side of Protein Regulation

项目总结 我的中心愿景是了解糖基化是如何作为一种早期营养感知机制发挥作用的 将细胞代谢状态与蛋白质功能联系起来。糖基化是一种进化上保守的非酶 蛋白质修饰，但其生物学意义尚未确定。广义地说，它描述了共价键 将葡萄糖和相关的糖衍生代谢物加到蛋白质上。此外，虽然前向反应 在没有酶的情况下，不同的脱糖酶FN3K和DJ-1催化这些所谓的“糖”的去除 加成物“。糖基化在生理学和医学上是一个未得到充分研究的领域，直到最近它还被认为是一种 与高血糖相关的被动标记物和非特异性蛋白损伤。相反，我最近的发现 与其他研究一起暗示，糖基化在营养信号和基因调控中扮演着更具活力的角色。 我一直在这些基础上开发一个研究项目，专注于了解生物 这一新的蛋白质标记的含义，特别是在糖代谢正常和异常的背景下。我 假设糖基化是一种古老的机制，它调节蛋白质的功能和细胞的行为 营养素的可利用性。解决这一假说的主要挑战是缺乏敏感的蛋白质组学。 研究这种蛋白质修饰的方法。在这里，我们将实现高分辨率的同位素标记和 基于亲和力浓缩的高级蛋白质组学策略首先获得对糖基化和 脱糖作用，包括它们的靶标偏好、化学计量比和反应动力学。接下来，我们将把它应用于 了解糖基化在不同代谢条件下差异调节蛋白质中的作用。在……里面 特别是，我们将重点研究我们预先排序的高优先级途径中的高度“可糖化”的蛋白质。 从我最近的低分辨率糖基化分析中出现，包括翻译起始和延伸 因子、代谢蛋白和组蛋白。在成功完成后，我们预计将确定i) FN3K和DJ-1敏感糖基化关键蛋白靶点的异同，II)早期区分， 中晚期糖基化和相应的脱糖靶标，包括蛋白质和特定氨基酸 在内部，iii)糖基化、葡萄糖内流和新陈代谢之间已建立的关系，以及iv)询问其 通过调节我们的“高优先途径”，在积极的代谢适应中发挥作用。这将具有重要的意义 医学意义，因为脱糖酶非常容易受到小分子的抑制。至 为此，我们的发现将奠定坚实的基础，并为开发新的 脱糖抑制剂。重要的是，除了有代谢和年龄相关疾病的临床应用外， 与糖尿病一样，脱糖疗法也可能在癌症治疗中占有一席之地。总之，我的实验室是独一无二的 准备好迎接这一挑战，这将在我们对代谢蛋白质的理解上有新的突破 调节并生成元数据，这些元数据将推动未来假说驱动和临床重要的糖基化 研究。

PROJECT SUMMARY My central vision is to understand how glycation functions as an early nutrient sensing mechanism that links cellular metabolic state to protein function. Glycation is an evolutionarily conserved non-enzymatic protein modification but its biological significance has not been established. Broadly, it describes the covalent addition of glucose and related sugar-derived metabolites on to proteins. Moreover, while the forward reaction is free of enzymes, distinct de-glycating enzymes FN3K and DJ-1 catalyze the removal of these so-called “sugar adducts”. Glycation is an under-served area in physiology and medicine and until recently it was perceived as a passive marker and non-specific protein damage associated with hyperglycemia. Conversely, my recent findings together with other studies allude to a more dynamic role of glycation in nutrient signaling and gene regulation. I have been building upon these to develop a research program focused on understanding the biological implications of this new protein mark, particularly in the context of normal and aberrant sugar metabolism. I hypothesize that glycation is an ancient mechanism that adjusts protein function and cell behavior in response to nutrient availability. The main challenge in addressing this hypothesis is the lack of sensitive proteomics approaches to study this protein modification. Here, we will implement a high-resolution isotope labeling and affinity enrichment based advanced proteomics strategy to first gain mechanistic insights into glycation and deglycation, including their target preference, stoichiometry, and reaction kinetics. Next, we will apply this to understand the role of glycation in differentially regulating proteins under distinct metabolic conditions. In particular, we will focus on highly “glycatable” proteins within our pre-ranked “high priority pathways” that emerged from my recent low-resolution glycation profiling and includes translation initiation and elongation factors, metabolic proteins, and histones. Upon successful completion, we anticipate to have i) identified similarities and differences in key protein targets of FN3K and DJ-1 sensitive glycation, ii) distinguished early, intermediate, and late glycation and corresponding deglycation targets, both proteins and specific amino acids within, iii) established relationship between glycation, glucose influx, and metabolism, and iv) interrogated its role in affirmative metabolic adaptation by regulating our “high priority pathways”. This will have significant medical implications because the de-glycating enzymes are highly amenable to small molecule inhibition. To that end, our findings will lay a strong foundation and provide the necessary scientific impetus to develop new inhibitors of deglycation. Importantly, beyond having clinical applications for metabolic and age-related disorders like diabetes, de-glycation therapies may also have a place in cancer treatment. In summary, my lab is uniquely equipped to undertake this challenge that will have break new grounds in our understanding of metabolic protein regulation and generate metadata that will drive future hypothesis-driven and clinically important glycation research.