GENETIC CONTROL OF PROTEOGLYCAN METABOLISM

蛋白多糖代谢的基因控制

• 批准号: 8011921
• 负责人: Jeffrey D Esko
• 金额: $ 10.2万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-05 至 2010-07-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8011921
• 关键词: Abbreviations; Acetylglucosamine; Address; Affect; Alleles; Atherosclerosis; Bacterial Artificial Chromosomes; Binding; Biochemical; Brain; Cells; Chinese Hamster; Chinese Hamster Ovary Cell; Chondroitin Sulfates; Cloning; Complementary DNA; Connective Tissue; Cytomegalovirus; Deacetylase; Enzymes; Family; Fibroblast Growth Factor; Fibroblast Growth Factor 2; GAG Gene; Gene Dosage; Gene Targeting; Genes; Genetic; Glucuronic Acids; Glutathione S-Transferase; Glycosaminoglycans; Goals; Green Fluorescent Proteins; Growth Disorders; Heparan Sulfate Biosynthesis; Heparan Sulfate Proteoglycan; Heparitin Sulfate; Hereditary Multiple Exostoses; Hexuronic Acids; Iduronic Acid; Immobilization; Individual; Isoenzymes; Lead; Ligands; Metabolism; Mus; Mutant Strains Mice; Neoplastic Cell Transformation; Oligosaccharides; Ovary; Pathway interactions; Physiological Processes; Plasmids; Process; Protein-Carbohydrate Interaction; Proteins; Proteoglycan; Regulation; Regulator Genes; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Screening procedure; Series; Specificity; Structure; Substrate Specificity; System; Tissues; Transferase; Vascular Endothelial Cell; Vascular Endothelial Growth Factors; human disease; insight; interest; member; mutant; novel strategies; polymerization; progesterone 11-hemisuccinate-(2-iodohistamine); programs; research study; sulfation; sulfotransferase

The long-range objective of this proposal is to gain insight into the mechanism of heparan sulfate proteoglycan assembly and its regulation. Heparan sulfate proteoglycans participate in a variety of physiological processes by binding, activation or immobilization of various protein ligands. These interactionsdepend to a large extent on the composition and fine structure of the heparan sulfate chains, which in turn depend on the substrate specificity of the various biosynthetic enzymes and regulatory factors. Thus, insight into the mechanismsthat cells use to synthesize and to regulate heparan sulfate composition may provide new avenues for developing agents to enhance or modulate protein- carbohydrate interactions. This information may lead to novel approaches for treating human diseases associated with altered heparan sulfate synthesis, such as neoplastic transformation, atherosclerosis, and various growth disorders associated with connective tissues. To achieve these long-range goals, we propose a series of genetic and biochemical experiments to probe the biosyntheticpathways of heparan sulfate synthesis in Chinese hamster ovary (CHO) cells and mice. Specifically,we plan to execute the following studies: 1. Identify genes affecting heparan sulfate biosynthesis. We will continueto isolate and characterize CHO cell mutants altered in glycosaminoglycansynthesis in order to characterize genes involved in chain initiation, polymerization and sulfation, with particularemphasis on heparan sulfate. In addition, we will search for regulatory genes in the system using multi-copy plasmidsto inactivatethe pathway through gene dosage effects. 2. Analyze the function of GIcA transferase-I in mice. W will target GlcAT-I in mice using the Cre-loxP system in order to ablate the gene in a tissue specific manner. Targeting the gene in this way will allow us to address whether a related isozyme (GlcAT-P) participatesin glycosaminoglycanformation in the brain. The mouse mutant would open up the possibility of studying the function of glycosaminoglycans in other tissues as well. 3. Determine the relationship between heparan sulfate copolymerase and pgsD. A controversy exists regarding the identity of pgsD and EXT, the putative heparan sulfate copolymerase. To resolve this problem, we will obtain the cDNA that encodes the pgsD locus by cloningthe gene from a stable correctant or by a sib-screening procedure. Characterizing various mutant alleles of pgsD that alter both GlcNAc and GIcA transferase activities or only the GlcA transferase may provide insight into structure and function of this interestinglocus. 4. Explore the specificity of the new NDST isozymcs in heparan sulfate processing. We will finish cloningthe fourth member of the N-deacetylase/N-sulfotransferase family and examine the substrate specificity of each isozyme by trapping and characterizing acceptor oligosaccharides. In additionwe will explore how the individual isozymes affects heparan sulfate biosynthesis in transfected cells.

该提案的长期目标是深入了解硫酸乙酰肝素蛋白多糖的组装机制 及其监管。硫酸乙酰肝素蛋白聚糖通过结合参与多种生理过程， 各种蛋白质配体的活化或固定。这些相互作用在很大程度上取决于成分 和硫酸乙酰肝素链的精细结构，这又取决于各种底物特异性 生物合成酶和调节因子。因此，深入了解细胞用于合成和分解的机制 调节硫酸乙酰肝素成分可能为开发增强或调节蛋白质的药物提供新途径 碳水化合物相互作用。这些信息可能会带来治疗与以下疾病相关的人类疾病的新方法： 硫酸乙酰肝素合成改变，例如肿瘤转化、动脉粥样硬化和各种生长障碍 与结缔组织有关。为了实现这些长期目标，我们提出了一系列遗传和生化方案 探究中国仓鼠卵巢（CHO）细胞硫酸乙酰肝素合成生物合成途径的实验 老鼠。具体来说，我们计划开展以下研究： 1. 鉴定影响硫酸乙酰肝素生物合成的基因。我们将继续分离和表征CHO细胞 改变糖胺聚糖合成的突变体，以表征参与链起始的基因， 聚合和硫酸化，特别强调硫酸乙酰肝素。此外，我们还将寻求监管 使用多拷贝质粒系统中的基因通过基因剂量效应灭活该途径。 2. 分析小鼠GlcA转移酶-I的功能。 W 将使用 Cre-loxP 系统靶向小鼠体内的 GlcAT-I 以便以组织特异性方式消除基因。以这种方式定位基因将使我们能够解决是否 相关同工酶（GlcAT-P）参与大脑中糖胺聚糖的形成。老鼠突变体会开放 研究糖胺聚糖在其他组织中的功能的可能性。 3.确定硫酸乙酰肝素共聚酶与pgsD之间的关系。对此存在争议 pgsD 和 EXT（假定的硫酸乙酰肝素共聚酶）的身份。为了解决这个问题，我们将获得cDNA 通过从稳定的校正子中克隆基因或通过同胞筛选程序来编码 pgsD 基因座。表征 pgsD 的各种突变等位基因改变 GlcNAc 和 GlcA 转移酶活性或仅改变 GlcA 转移酶可能 提供对这个有趣位点的结构和功能的深入了解。 4. 探索新NDST同工酶在硫酸乙酰肝素加工中的特异性。我们将完成克隆 N-脱乙酰酶/N-磺基转移酶家族的第四个成员，并通过以下方式检查每种同工酶的底物特异性 捕获和表征受体寡糖。此外，我们将探讨各个同工酶如何影响 转染细胞中硫酸乙酰肝素的生物合成。

项目成果

Hepatic heparan sulfate is a master regulator of hepcidin expression and iron homeostasis in human hepatocytes and mice.

硫酸肝素是人肝细胞和小鼠中铁调素表达和铁稳态的主要调节剂。

• 发表时间: 2019
• 作者: Poli, Maura;Anower;Asperti, Michela;Ruzzenenti, Paola;Gryzik, Magdalena;Denardo, Andrea;Gordts, Philip L S M;Arosio, Paolo;Esko, Jeffrey D
• 通讯作者: Esko, Jeffrey D

Biosynthesis of the linkage region of glycosaminoglycans: cloning and activity of galactosyltransferase II, the sixth member of the beta 1,3-galactosyltransferase family (beta 3GalT6).

糖胺聚糖连接区的生物合成：半乳糖基转移酶 II 的克隆和活性，半乳糖基转移酶 II 是 β 1,3-半乳糖基转移酶家族的第六个成员 (β 3GalT6)。

• 发表时间: 2001-12-21
• 作者: Bai, X;Zhou, D;Brown, J R;Crawford, B E;Hennet, T;Esko, J D
• 通讯作者: Esko, J D

Enzyme interactions in heparan sulfate biosynthesis: uronosyl 5-epimerase and 2-O-sulfotransferase interact in vivo.

硫酸乙酰肝素生物合成中的酶相互作用：糖醛酸基 5-差向异构酶和 2-O-磺基转移酶在体内相互作用。

• 发表时间: 2001-11-06
• 影响因子: 11.1
• 作者: Pinhal, M A;Smith, B;Olson, S;Aikawa, J;Kimata, K;Esko, J D
• 通讯作者: Esko, J D

Animal cell mutants defective in glycosaminoglycan biosynthesis.

• DOI: 10.1073/pnas.82.10.3197
• 发表时间: 1985-05-01
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: J. Esko;T. Stewart;W. H. Taylor
• 通讯作者: W. H. Taylor

CENTRAL ROLE FOR BROWN ADIPOSE TISSUE IN DYSLIPIDEMIA AND ATHEROSCLEROSIS DEVELOPMENT

棕色脂肪组织在血脂异常和动脉粥样硬化发展中的核心作用

• 发表时间: 2024-09-14
• 作者: M. Boon;J. F. Berbée;P. Khedoe;A. Bartelt;Linda P Brouwers;P. Gordts;J. Esko;P. Hiemstra;L. Havekes;L. Scheja;J. Heeren;P. Rensen
• 通讯作者: P. Rensen