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 项目概要 该研究计划的长期目标是阐明和理解所涉及的监管机制 哺乳动物细胞中糖胺聚糖 (GAG) 的生物合成。 GAG 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发挥关键作用，包括细胞信号传导和 发展。这些复杂的碳水化合物通常附着在核心蛋白上，称为蛋白聚糖， 位于细胞表面和细胞外基质中，链由交替组成 氨基葡萄糖和糖醛酸残基被异质 N- 和 O- 硫酸化。的生物合成 GAG 是一个非模板化过程，由位于酶的一大家族的协同活性驱动 高尔基体和内质网。硫酸化糖残基的排列和方向指定了 细胞表面上不同配体结合位点的位置，并且这些修饰可以在时间上变化 发育和跨组织空间。 GAG 结合配体的能力影响着基本的 细胞的特性、形成组织和器官的能力以及正常的生理机能。尽管关键功能 对于这些分子，关于引起这些分子的调节机制的知识存在显着差距 它们的可变组成和结合特性。通过多学科研究计划，利用 凭借功能基因组学、细胞生物学和糖生物学的优势，我们的目标是识别和表征 涉及控制细胞中 GAG 结构和功能的固有多样性的机制。特别是 有趣的是，染色质重塑复合物在发育过程中和发育过程中调节 GAG 组装的作用。 疾病状态，因为我们最近将多梳抑制复合体（PRC）的成员确定为新的 GAG 酶表达和组装的表观遗传修饰剂。我们假设定义了表观遗传和 转录程序调节不同细胞类型中生物合成酶的表达，从而调节其 与细胞外基质中的大量生长因子和其他结合伙伴相互作用。我们也瞄准 研究核心生物合成机制如何在内质网和高尔基体中进行原位调节。 我们计划探索内质网和内质网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调节分泌中糖基化的未知伴侣和/或支架蛋白 途径。为了开展这项工作，我们将利用我们在 GAG 结构、功能、 和调控以了解：（1）转录因子和染色质重塑复合物如何控制 GAG 生物合成酶的表达，影响 GAG 结构和功能，(2) 蛋白质-蛋白质如何 内质网和高尔基体之间的相互作用协调不同细胞类型中的蛋白聚糖组装，以及（3）如何 蛋白聚糖核心蛋白充当 GAG 组装的支架。总体而言，我们预计这一努力将显着 增进我们对控制糖基化的调控机制的了解并提供新的策略 以及操纵人类疾病中 GAG 生物发生的目标。