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: 9012950
• 负责人: THOMAS A GERKEN
• 金额: $ 15.74万
• 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 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9012950
• 关键词: Accounting; Address; Animal Model; Animals; Belief; Benign; Biological; Biological Process; Biology; C-terminal; Cardiovascular Diseases; Cardiovascular system; Cell Communication; Cell Culture Techniques; Cell Line; Cells; Complement; Data; Development; Disease; Embryonic Development; Enzymatic Biochemistry; Enzymes; Family; Fertility; Functional disorder; Glycopeptides; Goals; Health; Hormonal; Host Defense; Human; In Vitro; Individual; Inflammatory; Institution; Kinetics; Knock-out; Lead; Learning; Libraries; Link; Malignant Neoplasms; Membrane; Metabolic; Methods; Modification; Molecular; Mucins; Mus; Mutation; Online Systems; Pattern; Peptides; Play; Polypeptide N-acetylgalactosaminyltransferase; Polysaccharides; Prevalence; Process; Property; Protein Glycosylation; Protein Isoforms; Proteins; Regulation; Reporting; Reproduction; Role; Series; Site; Specificity; Structure; Substrate Specificity; Therapeutic; Tissues; Transferase; Work; animal tissue; base; design; fly; glycosylation; in vivo; inhibitor/antagonist; innovation; male; man; member; neoplastic; novel; novel strategies; polypeptide; ppGalNAc-T; predictive modeling; preference; public health relevance; sugar; therapeutic target; tool

 DESCRIPTION (provided by applicant): Most membrane and secreted proteins are decorated with mucin-type O-glycans which serve diverse biological roles. The mechanisms behind their biological roles and more specifically what governs O-glycan site selection and subsequent O-glycan elongation are largely unknown. The objective of this project is to elucidate the processes governing O-glycan site selection and O-glycan elongation at the enzyme and peptide substrate level in order to address the molecular mechanisms and biology of O-glycosylation. Mucin type O- glycosylation is vital to animal reproduction and development and increasingly linked to a wide range of rare to common disease states (such as hormonal/metabolic dysfunction, impaired host defense, inflammatory and cardiovascular diseases and even cancers) where it serves to modulate diverse biological functions including cell-cell interactions. Many disorders are linked to changes in expression or mutation of individual members of the large family (20 in man) of polypeptide-GalNAc transferases (ppGalNAc Ts) that initiate O-glycosylation by adding GalNAc to polypeptide Ser or Thr residues. O-glycosylation is absolutely required for embryonic development of the mouse and fly, relying on the Core 1 elongating transferase (T-synthase) in the mouse and several individual ppGalNAc T isoforms in the fly. How individual ppGalNAc T isoforms in this large family (or even T-synthase) can play such critical biological roles is unknown as the substrate specificity of these transferases have not been sufficiently characterized. That ppGalNAc T site selection is also modulated (positively or negatively) by prior glycosylation underscores its complexity and how much more needs to be learned of its specificity. The detailed characterization of ppGalNAc T isoform and elongating transferase specificity proposed in this project will lead to our understanding of the molecular mechanisms underlying the biological roles of O-glycosylation and will eventually lead to novel strategies to treat diseases of aberrant O- glycosylation. The AIMS of this project are to (1) expand the innovative use of a library of novel random peptide and glycopeptide substrates to fully characterize the specificity and basic enzymology of the transferases that initiate and elongate mucin type O-glycosylation, (2) to use these data to further develop sophisticated web based O-glycan predictive tools that include ppGalNAc T isoform and elongating transferase peptide and glycopeptide specificity, and (3) to develop a novel method for identifying in vivo isoform specific glycosylation targets using tissues from transferase knock-out animal models. These basic studies together with those of colleagues at other institutions will advance our understanding of the properties of these transferases, their targets (and resultant glycan structures) and ultimately the mechanisms of their biological role and function. These studies will significantly advance the field and will allw the development of novel specific inhibitors for potential use as targeted therapeutics.

 描述（由适用提供）：大多数膜和分泌的蛋白质都装饰有粘蛋白型O-聚糖，这些蛋白质具有多种生物学作用。它们的生物学作用背后的机制，更具体地说，是在很大程度上未知的O-Glycan位点选择和随后的O-聚糖伸长率的机制。该项目的目的是阐明在酶和胡椒底物水平上管理O-聚糖位点选择和O-聚糖伸长的过程，以解决O-糖基化的分子机制和生物学。粘蛋白型O-糖基化对于动物的繁殖和发育至关重要，并且越来越多地与普通疾病状态的广泛范围（例如，骑马/代谢功能障碍，宿主防御受损，炎症和心血管疾病，炎症和心血管疾病乃至癌症）至关重要。许多疾病都与多肽 - 盖尔纳克转移（PPGALNAC TS）的大家族单个成员的表达变化有关，这些疾病通过将galnac添加到多肽ser或thres或thres中。 O-糖基化是依赖于小鼠中的核1伸长转移（T-合酶）的胚胎发育和蝇的胚胎开发所必需的，并且果蝇中的几种单独的ppgalnac t同工型。这个大型家族（甚至T-合酶）中的单个PPGALNAC T同工型如何扮演这种关键的生物学作用是未知的，因为这些转移的底物特异性尚未得到充分表征。先前的糖基化也强调了其复杂性以及需要了解更多的特异性，也可以通过先前的糖基化来调节PPGALNAC T位点的选择。 PPGALNAC T同工型和伸长转移酶特异性的详细表征将导致我们对O-糖基化生物学作用的分子机制的理解，并最终导致治疗异常疾病的新型策略 O-糖基化。该项目的目的是（1）扩大新型随机肽和糖肽底物的创新使用，以充分表征转运的特异性和基本酶学，这些转移的特异性和基本酶学，这些转移和基本的酶学，并将粘液型粘液型O-糖基化型（2）用于进一步开发这些数据，以进一步开发基于复杂的web gygly IS extive intac ppgaln IS ETAC，并将其弹性;糖肽特异性和（3）开发一种新的方法，用于使用组织鉴定体内同工型特异性糖基化靶标 来自转移酶敲除动物模型。这些基本研究以及其他机构的同事的研究将提高我们对这些转移的特性，它们的目标（以及由此产生的聚糖结构）以及最终其生物学作用和功能机制的理解。这些研究将大大推进该领域，并将全力发展新型特异性抑制剂作为靶向治疗的潜在用途。