Investigating the molecular mechanisms of glycosaminoglycan assembly

研究糖胺聚糖组装的分子机制

• 批准号: 10715380
• 负责人: Ryan Joseph Weiss
• 金额: $ 37.75万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-15 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10715380
• 关键词: Anabolism; Animals; Binding; Binding Sites; Biochemical; Biogenesis; Carbohydrates; Cell physiology; Cell surface; Cells; Cellular biology; Chromatin Remodeling Factor; Complex; Core Protein; Development; Disease; Endoplasmic Reticulum; Enzymes; Epigenetic Process; Extracellular Matrix; Family; Genetic Transcription; Glucosamine; Glycobiology; Glycosaminoglycans; Goals; Golgi Apparatus; Growth Factor; Homeostasis; Human; In Situ; Infection; Interdisciplinary Study; Knowledge; Ligand Binding; Location; Malignant Neoplasms; Mammalian Cell; Modification; Molecular; Molecular Chaperones; Organ; Pathway interactions; Physiology; Play; Polycomb; Polysaccharides; Process; Property; Proteoglycan; Proteomics; Regulation; Research; Role; Scaffolding Protein; Signal Transduction; Specific qualifier value; Structure; Sugar Acids; Sulfate; Techniques; Tissues; Uronic Acids; Work; cell type; developmental disease; functional genomics; glycosylation; human disease; interest; member; new therapeutic target; novel; pathogen; programs; protein protein interaction; proteoglycan core protein; scaffold; sugar; transcription factor; virtual

Title: Investigating the molecular mechanisms of glycosaminoglycan assembly Project Summary The long-term goal of this research program is to elucidate and understand the regulatory mechanisms involved in the biosynthesis of glycosaminoglycans (GAGs) in mammalian cells. GAGs are long, linear polysaccharides that are expressed on all animal cells and play a key role in many cellular processes, including cell signaling and development. These complex carbohydrates are typically attached to core proteins, known as proteoglycans, located on the cell surface and in the extracellular matrix, and the chains are composed of alternating glucosamine and uronic acid sugar residues that are heterogeneously N- and O-sulfated. The biosynthesis of GAGs is a non-templated process, driven by the concerted activity of a large family of enzymes localized to the Golgi and endoplasmic reticulum. The arrangement and orientation of the sulfated sugar residues specify the location of distinct ligand binding sites on the cell surface, and these modifications can vary temporally during development and spatially across tissues. The capacity of GAGs to bind ligands impacts the fundamental properties of cells, the ability to form tissues and organs, and normal physiology. Despite the key functions of these molecules, there is a significant gap in knowledge regarding the regulatory mechanisms that give rise to their variable composition and binding properties. Through a multidisciplinary research program that leverages strengths in functional genomics, cell biology, and glycobiology, we aim to identify and characterize the mechanisms involved in controlling the inherent diversity of GAG structure and function in cells. Of particular interest, is the role of chromatin remodeling complexes in regulating GAG assembly during development and in disease states, as we recently identified members of the polycomb repressive complex (PRC) as novel epigenetic modifiers of GAG enzyme expression and assembly. We hypothesize that defined epigenetic and transcriptional programs tune the expression of biosynthetic enzymes in distinct cell types, which modulates their interaction with a plethora of growth factors and other binding partners in the extracellular matrix. We also aim to investigate how the core biosynthetic machinery is regulated in situ in the endoplasmic reticulum and Golgi. We plan to explore the physical association of the biosynthetic enzymes and core proteoglycans in the ER and Golgi and identify unknown chaperone and/or scaffolding proteins that may tune glycosylation in the secretory pathway. To carry out this work, we will leverage our historic strengths in the analysis of GAG structure, function, and regulation to understand: (1) how transcription factors and chromatin remodeling complexes control the expression of GAG biosynthetic enzymes, which impacts GAG structure and function, (2) how protein-protein interactions in the ER and Golgi orchestrate proteoglycan assembly in distinct cell types, and (3) how proteoglycan core proteins act as scaffolds for GAG assembly. Overall, we expect this endeavor to significantly advance our knowledge regarding the regulatory mechanisms controlling glycosylation and offer new strategies and targets to manipulate GAG biogenesis in human disease.

标题：研究糖胺聚糖组装的分子机制 项目摘要 该研究计划的长期目标是阐明和了解涉及的监管机制 在哺乳动物细胞中糖胺聚糖（GAGS）的生物合成中。插科打术是长的线性多糖 在所有动物细胞上表达并在许多细胞过程中起关键作用，包括细胞信号和 发展。这些复杂的碳水化合物通常连接到核心蛋白（称为蛋白聚糖）上 位于细胞表面和细胞外基质中，链由交替组成 葡萄糖胺和尿酸糖残基是异构N-和O硫化的。生物合成 插科打s是一个非模范过程，是由局部属于该酶的大型酶的一致活动驱动的 高尔基和内质网。硫酸糖残基的排列和方向指定 不同配体结合位点在细胞表面的位置，这些修饰在时间上可能会变化 开发和空间跨组织。插科打结合配体的能力会影响基本 细胞的特性，形成组织和器官的能力以及正常的生理学。尽管有关键功能 这些分子，关于导致调节机制的知识存在很大的差距 它们的可变组成和结合特性。通过利用的多学科研究计划 功能基因组学，细胞生物学和糖生物学方面的优势，我们旨在识别和表征 控制细胞中GAG结构和功能的固有多样性涉及的机制。特别 兴趣，是染色质重塑复合物在调节开发过程和在调节插孔组件中的作用 疾病状态，正如我们最近确定的Polycomb抑制复合物（PRC）成员是新颖的 GAG酶表达和组装的表观遗传修饰符。我们假设定义的表观遗传学和 转录程序在不同的细胞类型中调整生物合成酶的表达，这使其调节它们 在细胞外基质中与大量生长因子和其他结合伴侣的相互作用。我们也瞄准 为了研究如何在内质网和高尔基体中原位调节核心生物合成机械。 我们计划探索ER中生物合成酶和核心蛋白聚糖的物理关联 高尔基体并识别可能在分泌中调节糖基化的未知伴侣和/或脚手架蛋白 路径。为了进行这项工作，我们将利用我们的历史优势在分析插科打结构，功能， 并了解：（1）转录因子和染色质重塑配合物如何控制 GAG生物合成酶的表达，会影响GAG结构和功能，（2）蛋白质蛋白如何 ER和高尔基体中的相互作用以不同的细胞类型编排蛋白聚糖组装，以及（3） 蛋白聚糖核心蛋白充当插科打组装的支架。总的来说，我们希望这项工作能够大大 提高我们有关控制糖基化的调节机制的知识，并提供新的策略 和操纵人类疾病中堵嘴生物发生的靶标。