In vitro synthesis of recombinant heparan sulfate

重组硫酸乙酰肝素的体外合成

• 批准号: 8038366
• 负责人: JIAN LIU
• 金额: $ 40.77万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-13 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8038366
• 关键词: Abbreviations; Adverse effects; Anticoagulants; Antineoplastic Agents; Antithrombins; Antiviral Agents; Binding; Biological; Biological Process; Blood coagulation; Cell Differentiation process; Cell surface; Deacetylation; Development; Disaccharides; Enzymes; Escherichia coli; Extracellular Matrix; Glucosamine; Heparan Sulfate Biosynthesis; Heparin; Heparitin Sulfate; Iduronic Acid; In Vitro; Inflammatory Response; Inorganic Sulfates; Lead; Libraries; Malignant Neoplasms; Methods; Modification; Oligosaccharides; Pasteurella; Pattern; Physiological; Polysaccharides; Positioning Attribute; Predisposition; Reagent; Recombinants; Series; Signal Transduction; Specificity; Structure; Structure-Activity Relationship; Substrate Specificity; Taiwan; Therapeutic; Therapeutic Agents; Transferase; UDP-glucosamine; Universities; Unspecified or Sulfate Ion Sulfates; base; chemical synthesis; design; epimerase; epimerization; heparosan synthase; improved; interest; novel; novel strategies; pathogen; public health relevance; success; sulfation; sulfotransferase

DESCRIPTION (provided by applicant): Heparan sulfate (HS) is an essential glycan that is present in large quantities on the cell surface and in the extracellular matrix. HS participates in a variety of physiological and pathophysiological functions, including blood coagulation, inflammatory response and cell differentiation. HS is a highly sulfated polysaccharide. Heparin, a special form of HS, is a commonly used anticoagulant drug. The wide range of biological functions of HS attracts considerable interest in exploiting heparin or heparin-like molecules for the development of anticancer and antiviral drugs. The uniquely distributed sulfation pattern of HS is believed to regulate its functional specificity. However, chemical synthesis of HS, especially those larger than hexasaccharides, is extremely difficult. Using HS biosynthetic enzymes, our labs can produce an array of HS with unique sulfation patterns and functions. Our success has proved the feasibility of conducting enzyme-based synthesis of HS with unique biological activities. The long term aim of this project is to develop a method to synthesize HS with high structural precision. Our hypothesis is that the distribution of N- sulfoglucosamine residues determines the susceptibility of all subsequent modifications during heparan sulfate biosynthesis, including epimerization, 2-O-sulfation, 6-O- sulfation, and 3-O-sulfation. In this proposal, we will synthesize structurally defined oligosaccharides carrying N-sulfoglucosamine residues using glycosyl transferases. We will then determine the substrate specificities of a variety of HS biosynthetic enzymes with the aim of precisely positioning the 6-O-sulfo and 3-O-sulfo groups as well as 2-O- sulfo iduronic acid within oligosaccharides. Finally, we plan to utilize this method to investigate novel anticoagulant HS structures. The success of this project will lead a comprehensive new approach to investigate the structure and function relationship of HS, potentially leading to the development of novel HS-based therapeutic reagents. Public Health Relevance: Heparan sulfate (HS) is a highly sulfated polysaccharide. Heparin, a special form of HS, is a commonly used anticoagulant drug. The wide range of biological functions of HS attracts considerable interest in exploiting heparin or heparin-like molecules for the development of anticancer and antiviral drugs. The uniquely distributed sulfation pattern of HS is believed to regulate its functional specificity. The long term aim of this project is to develop a method to synthesize HS with high structural precision. The success of this project will lead a comprehensive new approach to investigate the structure and function relationship of HS, potentially leading to the development of novel HS-based therapeutic reagents.

描述（由申请人提供）：硫酸乙酰肝素（HS）是一种必需聚糖，大量存在于细胞表面和细胞外基质中。 HS参与多种生理和病理生理功能，包括凝血、炎症反应和细胞分化。 HS是一种高度硫酸化的多糖。肝素是 HS 的一种特殊形式，是常用的抗凝药物。 HS 广泛的生物学功能引起了人们对利用肝素或类肝素分子开发抗癌和抗病毒药物的极大兴趣。 HS 独特分布的硫酸化模式被认为调节其功能特异性。然而，HS，特别是那些大于六糖的化学合成极其困难。使用 HS 生物合成酶，我们的实验室可以生产一系列具有独特硫酸化模式和功能的 HS。我们的成功证明了酶法合成具有独特生物活性的HS的可行性。该项目的长期目标是开发一种高结构精度合成HS的方法。我们的假设是，N-磺基葡萄糖胺残基的分布决定了硫酸乙酰肝素生物合成过程中所有后续修饰的敏感性，包括差向异构化、2-O-硫酸化、6-O-硫酸化和3-O-硫酸化。在本提案中，我们将使用糖基转移酶合成携带 N-磺基葡萄糖胺残基的结构明确的寡糖。然后，我们将确定各种 HS 生物合成酶的底物特异性，目的是在寡糖中精确定位 6-O-磺基和 3-O-磺基以及 2-O-磺基艾杜糖醛酸。最后，我们计划利用这种方法来研究新型抗凝剂 HS 结构。该项目的成功将带来一种全面的新方法来研究 HS 的结构和功能关系，并有可能导致新型基于 HS 的治疗试剂的开发。公共健康相关性：硫酸乙酰肝素 (HS) 是一种高度硫酸化的多糖。肝素是 HS 的一种特殊形式，是常用的抗凝药物。 HS 广泛的生物学功能引起了人们对利用肝素或类肝素分子开发抗癌和抗病毒药物的极大兴趣。 HS 独特分布的硫酸化模式被认为调节其功能特异性。该项目的长期目标是开发一种高结构精度合成HS的方法。该项目的成功将带来一种全面的新方法来研究 HS 的结构和功能关系，并有可能导致新型基于 HS 的治疗试剂的开发。