3-O-sulfation of heparan sulfate as a regular of protein function

硫酸乙酰肝素的 3-O-硫酸化作为蛋白质功能的调节

• 批准号: 10400697
• 负责人: Geert-Jan Boons
• 金额: $ 45.17万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10400697
• 关键词: Address; Anticoagulants; Binding; Binding Proteins; Biological; Biological Process; Biology; Blood Coagulation Factor; Blood Vessels; Blood coagulation; Carbohydrates; Cell Adhesion; Cell surface; Cells; Chemicals; Code; Collection; Complement Activation; Complex; Data; Deep Vein Thrombosis; Disaccharides; Disease; Endothelial Cells; Endothelium; Engineering; Enzymatic Biochemistry; Enzymes; Epitopes; Extracellular Matrix; Gene Family; Goals; Growth Factor; Heparin; Heparitin Sulfate; Human; Infection; Inflammation; Inflammatory; Isoenzymes; Lead; Libraries; Life; Ligands; Malignant Neoplasms; Mammalian Cell; Mediating; Methodology; Methods; Modality; Modification; Nerve Degeneration; Oligosaccharides; Polysaccharides; Protein Isoforms; Proteins; Pulmonary Embolism; Recombinants; Signal Transduction; Signaling Protein; Site; Specificity; Structure; Structure-Activity Relationship; Substrate Specificity; Sulfate; Surface; Technology; Therapeutic; Thromboembolism; Vascular Diseases; Vertebral column; cell type; chemokine; cytokine; gain of function; human disease; interest; mast cell; morphogens; pathogen; prevent; programs; protein function; recruit; sulfotransferase; therapeutic lead compound

Project summary/abstract Heparin is a highly sulfated carbohydrate produced by mast cells that is widely used as an anticoagulant to prevent deep vein thrombosis, pulmonary embolism and arterial thromboembolism. Heparan sulfate (HS) is structurally related to heparin and decorates the surface of all most all human cells. It interacts with a multitude of proteins including chemokines and cytokines; blood coagulation factors; growth factors and morphogens, proteins involved in complement activation, and cell adhesion and signaling proteins. HS has been implicated in human diseases such as vascular diseases, inflammation, infections and neurodegeneration. It has been postulated that HS encodes information by an ability to recruit proteins in a context-dependent manner. Although the “HS-sulfate code” hypothesis is conceptually appealing, little is known about the ligand specificities of HS binding proteins and the substrate specificities of HS-biosynthetic enzymes. As a result, it is not known whether isoforms of HS biosynthetic enzymes can create distinctive epitopes that in a cellular context can recruit specific HS- binding proteins. An understanding of the specificities of HS binding also offers opportunities to develop therapeutic modalities. In this program, chemical and chemo-enzymatic methodologies will be developed that will make it possible to prepare a focused library of biologically relevant HS epitopes with and without a 3-O-sulfate moiety. These compounds will be used to develop an HS-microarray to establish ligand requirement of HS-binding proteins. Compounds of interest will be further explored for their ability to interfere in the binding and cellular activation of endothelial cells. We will focus on HS-binding proteins involved in blood coagulation, inflammation and vascular disease. The established structure-activity relationships are expected to provide therapeutic lead compounds. The HS-microarray will also be used as a general platform to discover and characterize HS-binding proteins requiring a 3-O-sulfate. In addition, the synthetic compounds will be used to define substrate requirements of the various 3-O-sulfotransferases. Collectively, our studies will address the question of whether isoforms of 3-O-sulfotransferases can create distinctive epitopes that can recruit specific HS-binding proteins, to mediate various biological processes. 1

项目概要/摘要 肝素是一种由肥大细胞产生的高度硫酸化的碳水化合物，被广泛用作抗凝剂 预防深静脉血栓、肺栓塞和动脉血栓栓塞。硫酸乙酰肝素 (HS) 在结构上与肝素相关，并装饰几乎所有人类细胞的表面。它相互作用 含有多种蛋白质，包括趋化因子和细胞因子；凝血因子；生长 因子和形态发生素、参与补体激活的蛋白质以及细胞粘附和信号传导 蛋白质。 HS 与人类疾病有关，如血管疾病、炎症、 感染和神经变性。据推测，HS 通过以下能力对信息进行编码： 以上下文相关的方式招募蛋白质。尽管“HS-硫酸盐代码”假说是 从概念上讲很有吸引力，但人们对 HS 结合蛋白的配体特异性和 HS-生物合成酶的底物特异性。因此，尚不清楚 HS 的亚型是否 生物合成酶可以产生独特的表位，在细胞环境中可以招募特定的 HS- 结合蛋白。了解 HS 结合的特殊性也提供了发展的机会 治疗方式。 在该计划中，将开发化学和化学酶方法，使其成为可能 可以制备含有或不含 3-O-硫酸盐的生物学相关 HS 表位的集中文库 部分。这些化合物将用于开发 HS 微阵列以确定配体需求 HS 结合蛋白。将进一步探索感兴趣的化合物干扰的能力 内皮细胞的结合和细胞活化。我们将重点关注血液中涉及的 HS 结合蛋白 凝血、炎症和血管疾病。建立的构效关系为 有望提供治疗性先导化合物。 HS 微阵列也将用作通用 发现和表征需要 3-O-硫酸盐的 HS 结合蛋白的平台。此外， 合成化合物将用于确定各种 3-O-磺基转移酶的底物要求。 总的来说，我们的研究将解决 3-O-磺基转移酶亚型是否可以 创建独特的表位，可以招募特定的 HS 结合蛋白，以介导各种生物 流程。 1