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是如何作为一种早期营养感应机制发挥作用的， 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添加到蛋白质上。此外，虽然前向反应 不含酶，不同的去糖化酶FN3K和DJ-1催化去除这些所谓的"糖 加合物"。糖基化是生理学和医学中一个服务不足的领域，直到最近，它被认为是一个 被动标记物和与高血糖症相关的非特异性蛋白质损伤。相反，我最近的发现 与其他研究一起暗示了糖化在营养信号和基因调控中的更动态作用。 我一直在这些基础上开发一个研究计划，重点是了解生物学 这种新的蛋白质标记的意义，特别是在正常和异常糖代谢的背景下。我 假设糖基化是一种古老机制，它调节蛋白质功能和细胞行为， to nutrition营养availability可用性.解决这一假设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，我们将实施高分辨率同位素标记， 基于亲和富集的先进蛋白质组学策略，首先获得对糖化和 去糖基化，包括它们的目标偏好、化学计量和反应动力学。接下来，我们将把它应用到 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 特别是，我们将集中在我们预先排序的"高优先级途径"中的高度"糖基化"蛋白质， 从我最近的低分辨率糖化分析中出现，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，我们预计将有i）确定 FN3K和DJ-1敏感性糖基化的关键蛋白质靶标的相似性和差异，ii）早期区分， 中间和晚期糖基化以及相应的去糖基化靶标，包括蛋白质和特定氨基酸 iii）确定糖化、葡萄糖内流和代谢之间的关系，以及iv）询问其 通过调节我们的“高优先级途径”，在积极的代谢适应中发挥作用。这将对 医学意义，因为去糖化酶非常适合小分子抑制。到 为此，我们的研究结果将奠定坚实的基础，并为开发新产品提供必要的科学动力 去糖化抑制剂。重要的是，除了对代谢和年龄相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。总之，我的实验室是独一无二的 我们有能力接受这一挑战，这将为我们对代谢蛋白质的理解开辟新的领域。 调节并生成元数据，以驱动未来假设驱动的临床重要糖化 research.