Understand Glycation: The Sweet Side of Protein Regulation

了解糖化：蛋白质调节的甜蜜一面

• 批准号: 10818932
• 负责人: viraj sanghvi
• 金额: $ 41.13万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10818932
• 关键词: Address; Affinity; Amino Acids; Biological; Clinical; Diabetes Mellitus; Disease; Elongation Factor; Enzymes; Excision; Foundations; Future; Gene Expression Regulation; Genes; Glucose; Histones; Hyperglycemia; Isotope Labeling; Kinetics; Link; Malignant Neoplasms; Medical; Medicine; Metabolic; Metabolic Diseases; Metabolism; Metadata; Nutrient; Nutrient availability; PARK7 gene; Pathway interactions; Peptide Initiation Factors; Physiology; Post-Translational Protein Processing; Proteins; Proteomics; Reaction; Regulation; Research; Resolution; Role; Side; Signal Transduction; Vision; adduct; age related; cancer therapy; cell behavior; clinical application; detection of nutrient; glycation; inhibitor; insight; preference; programs; protein function; response; small molecule; stoichiometry; sugar; underserved area

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.

项目摘要 我的中心愿景是了解糖基化如何作为早期营养感应机制的作用 将细胞代谢状态与蛋白质功能联系起来。糖基是一种进化配置的非酶 蛋白质修饰，但其生物学意义尚未确定。从广义上讲，它描述了共价 将葡萄糖和相关糖衍生的代谢产物添加到蛋白质上。而且，前进反应 不含酶，独特的去糖化酶FN3K和DJ-1催化了这些所谓的“糖”的去除 加合物”。糖化是生理和医学的服务不足的领域，直到最近它被认为是一种 被动标记和非特异性蛋白质损伤与高血糖有关。相反，我最近的发现 与其他研究一起，暗示了糖基化在营养信号传导和基因调节中的动态作用。 我一直在基础上建立一个研究计划，以了解生物学 这种新蛋白质标记的含义，特别是在正常和异常的糖代谢的背景下。我 假设糖基化是一种古老的机制，可在反应中调节蛋白质功能和细胞行为 养分可用性。解决这一假设的主要挑战是缺乏敏感蛋白质组学 研究这种蛋白质修饰的方法。在这里，我们将实施高分辨率的同位素标签，并 基于亲和力富集的先进蛋白质组学策略，以首先获得机械洞察力，并 脱胶，包括其目标偏好，化学计量和反应动力学。接下来，我们将其应用于 了解在不同的代谢条件下糖化在不同调节的蛋白质中的作用。在 特别是，我们将专注于在我们的预先排名的“高优先途径”中高度“可糖化”蛋白 从我最近的低分辨率糖基谱分析中出现，包括翻译计划和伸长率 因素，代谢蛋白和组蛋白。成功完成后，我们预计会有i）确定 FN3K和DJ-1敏感糖基化的关键蛋白质靶标的相似性和差异，ii）早期区分， 蛋白质和特异性氨基酸都中间糖化和相应的脱糖化靶标 iii）在糖化，葡萄糖影响和代谢之间建立了关系，iv）审问了它 通过调节我们的“高优先途径”，在肯定代谢适应中的作用。这将具有重要的 医学意义是因为去糖化酶高度适合小分子抑制作用。到 这一目的，我们的发现将奠定坚实的基础，并提供必要的科学动力来发展新的基础 程度抑制剂。重要的是，除了对代谢和年龄有关疾病的临床应用外 像糖尿病一样，去糖化疗法也可能在癌症治疗中占有一席之地。总而言之，我的实验室是独特的 能够承担这一挑战，这将在我们对代谢蛋白的理解中构成新的理由 调节和产生元数据，将推动未来假设驱动和临床上重要的糖化 研究。