Initiation and Regulation of Mucin-Type O-Glycosylation

粘蛋白型 O-糖基化的启动和调节

• 批准号: 10118475
• 负责人: THOMAS A GERKEN
• 金额: $ 33.81万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10118475
• 关键词: Acetylgalactosamine; Base Sequence; Biochemical; Biological; Biological Process; Cardiovascular Diseases; Cardiovascular system; Cell Communication; Cell surface; Cells; Charge; Colon; Complex; Coronary Arteriosclerosis; Crystallization; Data; Defect; Development; Disease; Drug Industry; Embryonic Development; Enzymes; Family; Fertility; Funding; Future; Glycopeptides; Glycoproteins; Goals; Hormonal; Host Defense; Human; Immune response; In Vitro; Individual; Inflammation; Inflammatory; Kinetics; Knowledge; Link; Malignant Neoplasms; Membrane; Metabolic; Methods; Modeling; Modification; Molecular; Mucins; Mus; Online Systems; Organism; Pathway interactions; Pattern; Peptides; Play; Polypeptide N-acetylgalactosaminyltransferase; Polysaccharides; Post-Translational Protein Processing; Prevalence; Process; Property; Protein Glycosylation; Protein Isoforms; Proteins; Regulation; Renal function; Research; Role; Series; Site; Specificity; Structure; Substrate Specificity; Testing; Therapeutic; Therapeutic Uses; Transferase; Tumor Markers; Work; base; fly; genome wide association study; glycoproteomics; glycosylation; glycosyltransferase; in vivo; man; neoplastic; novel; polypeptide; protein aminoacid sequence; protein degradation; receptor; sialylation; sugar; tool

Mucin-type protein O-glycosylation (henceforth called O-glycosylation) is a ubiquitous and essential post translational modification of higher organisms. Most proteins passing through the secretory pathway are decorated with a wide range of mucin-type O-glycans which serve diverse biological functions. Hence, many biological processes and disease states are linked to normal or abnormal O-glycosylation including coronary artery disease, the regulation of kidney function, organogenisis, embryonic development, multiple cancers and fertility. Importantly, the loss of single O-glycan initiating and elongating transferases is developmentally lethal in the fly and mouse, respectively. Presently it is not well understood how these transferases chose their specific targets and what features of their substrates modulate their activities. Such an understanding, at the molecular level, is necessary for deducing the biological roles of O-glycosylation and for predicting sites of O- glycosylation. By understanding all of the factors involved in substrate selection new avenues will open for the development of novel and selective strategies to treat diseases of aberrant O-glycosylation including cancers. Furthermore, the ability to predict transferase specific sites of O-glycosylation will be invaluable for the interpretation of O-glycoproteomics data and for identifying the targets of glycosyltransferases linked to disease from genome wide association studies (GWAS). The planned research will focus on the large family of GalNAc-Ts (T1-T20) that initiate O-glycosylation and the core transferases (C1GALT1, B3GNT6, GCNT1 and ST6GalNAc-1 & 2) that perform the first step(s) of O-glycan elongation. This research aims to characterize the unique peptide and glycopeptide substrate specificities of the GalNAc-Ts as well as to identify additional substrate features such as clustered charges and prior glycosylation that may control O- glycosylation. Our major working hypothesis is that O-glycan site selection and specific elongation are modulated by the properties of the peptide with one component being the charge distribution of residues flanking the acceptor site. This work will provide an unprecedented understanding of GalNAc-T substrate selection, achieved by correlating our specificity and kinetics data with the crystal structures of substrates bound or modeled onto the GalNAc-Ts and the core elongating transferases. Additional studies will involve characterizing the role of prior Thr versus Ser O-glycosylation, the glycosylation of Tyr residues and the further refinement of our web based isoform specific O-glycosylation prediction tool ISOGlyP. Together, these basic studies will greatly advance our understanding of the properties of these transferases and how they chose their targets and ultimately the mechanisms of their biological role and function in disease with an eventual goal to develop useful therapeutics for the treatment of diseases of aberrant O-glycosylation.

粘蛋白型蛋白O-糖基化（以下简称O-糖基化）是一种普遍存在的必需的后处理过程 高等生物的翻译修饰。大多数通过分泌途径的蛋白质是 用广泛的粘蛋白型O-聚糖装饰，其具有多种生物学功能。所以很多 生物学过程和疾病状态与正常或异常的O-糖基化有关，包括冠状动脉 动脉疾病、肾功能调节、器官发生、胚胎发育、多种癌症和 生育重要的是，单个O-聚糖起始和延伸转移酶的缺失在发育上是致命的 分别在苍蝇和老鼠身上。目前还不清楚这些转移酶如何选择它们的活性。 特定的目标和什么样的功能，他们的底物调节他们的活动。这样的理解，在 在分子水平上，对于推断O-糖基化的生物学作用和预测O-糖基化位点是必要的。 糖基化通过了解底物选择中涉及的所有因素，将为底物选择开辟新的途径。 开发新的和选择性的策略来治疗异常O-糖基化的疾病，包括癌症。 此外，预测0-糖基化的转移酶特异性位点的能力对于本领域技术人员来说将是无价的。 O-糖蛋白组学数据的解释和用于鉴定与糖基转移酶相关的靶点。 全基因组关联研究（GWAS）计划中的研究将集中在大家庭 启动O-糖基化的GalNAc-Ts（T1-T20）和核心转移酶（C1 GALT 1、B3 GNT 6、GCNT 1 和ST 6 GalNAc-1和2），其执行O-聚糖延伸的第一步。本研究旨在 表征GalNAc-Ts的独特肽和糖肽底物特异性以及鉴定 额外的底物特征，如成簇电荷和先前的糖基化， 糖基化我们的主要工作假设是，O-聚糖位点选择和特异性延伸是 由肽的性质调节，其中一个组分是残基的电荷分布 位于受体位点的侧翼。这项工作将提供一个前所未有的了解GalNAc-T底物 选择，通过将我们的特异性和动力学数据与底物的晶体结构相关联来实现 结合或模仿GalNAc-Ts和核心延伸转移酶。其他研究将涉及 表征了先前Thr相对于Ser 0-糖基化、Tyr残基的糖基化和进一步的糖基化的作用， 我们的基于网络的异构体特异性O-糖基化预测工具ISOGlyP的改进。这些基本的 这些研究将极大地促进我们对这些转移酶的性质以及它们如何选择它们的 目标和最终的机制，其生物学作用和功能的疾病，最终目标是 开发用于治疗异常O-糖基化疾病的有用治疗剂。