Enzymatic Degradation of Glycosaminoglycans

糖胺聚糖的酶降解

• 批准号: 8008961
• 负责人: RAM SASISEKHARAN
• 金额: $ 2.7万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-25 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8008961
• 关键词: Address; Area; Biochemical; Bioinformatics; Biological; Biological Process; Cells; Chemicals; Chondroitinases; Complex; Data; Data Set; Databases; Development; Environment; Enzymatic Biochemistry; Eukaryotic Cell; Extracellular Matrix; GAG Gene; Glycosaminoglycan Degradation Pathway; Glycosaminoglycans; Goals; Grant; Heart; Heterogeneity; Hyaluronidase; Injury; International; Low-Molecular-Weight Heparin; Modeling; Multiple Myeloma; Nature; Organ; Phenotype; Polysaccharides; Proteins; Recovery of Function; Research; Research Personnel; Role; Spinal Cord; Structure; Structure-Activity Relationship; Techniques; Technology; Time; Tissues; analytical tool; chondroitin sulfate glycosaminoglycan; depolymerization; experience; extracellular; new therapeutic target; novel; novel strategies; programs; relational database; tool

DESCRIPTION (provided by applicant): Glycosaminoglycans (GAGs) are complex polysaccharides found in abundance at the cell-extracellular matrix (ECM) interface of eukaryotic cells. There has been a paradigm shift in understanding the importance of the dynamic nature of the cell-ECM interactions (historically the ECM was considered an inert material that hydrated the cells) in influencing phenotype at cellular and higher order tissue and organ levels. At the heart of this paradigm are the specific interactions between the chemically heterogeneous GAGs and numerous proteins in the extracellular environment. Thus, understanding the structure-function relationship of GAGs has gained importance both as a fundamental field and for its potential in identification of novel therapeutic targets. However, progress towards this goal has been limited in the past due to the polydispersity and chemical heterogeneity of GAGs that posed numerous challenges for their isolation and characterization. For several years, we have focused our efforts in addressing these challenges to develop enzymatic tools for predictable depolymerization of GAGs to decode their sequence information as well as sensitive analytical techniques to accurately detect and characterize extremely small amounts of GAGs available from cells or tissues. Focusing on the 2 important classes of GAGs viz. HSGAGs and CSGAGs we have successfully demonstrated the utility of our tools to develop rapid and robust technologies for sequencing GAGs. In this study we seek to expand on the development of our tools to focus on biochemical and biological aspects of HSGAG and CSGAG structure-function relationships. Finally, we seek to develop a database platform to capture and disseminate information and data pertaining to GAG structure function relationships which is a novel approach in the context of the GAG research area. We believe that our study would truly accelerate the progress of understanding GAG-structure-function relationships in fundamental biological processes-a research area that is gaining increasing importance.

描述（由申请人提供）：糖胺聚糖（GAG）是在真核细胞的细胞-细胞外基质（ECM）界面大量发现的复杂多糖。在理解细胞-ECM 相互作用的动态性质（历史上 ECM 被认为是水合细胞的惰性材料）在影响细胞和更高阶组织和器官水平的表型方面的重要性方面，已经发生了范式转变。该范例的核心是化学异质性 GAG 与细胞外环境中众多蛋白质之间的特定相互作用。因此，了解 GAG 的结构与功能关系作为一个基础领域及其在识别新治疗靶点方面的潜力已变得越来越重要。然而，由于 GAG 的多分散性和化学异质性，给其分离和表征带来了许多挑战，过去实现这一目标的进展受到限制。多年来，我们一直致力于解决这些挑战，开发可预测的 GAG 解聚酶工具，以解码其序列信息，以及灵敏的分析技术，以准确检测和表征细胞或组织中极少量的 GAG。重点关注 2 类重要的 GAG，即。 HSGAG 和 CSGAG 我们已成功展示了我们的工具在开发快速、稳健的 GAG 测序技术方面的实用性。在这项研究中，我们寻求扩展我们工具的开发，重点关注 HSGAG 和 CSGAG 结构功能关系的生化和生物学方面。最后，我们寻求开发一个数据库平台来捕获和传播与 GAG 结构功能关系相关的信息和数据，这是 GAG 研究领域的一种新颖方法。我们相信，我们的研究将真正加速理解基本生物过程中 GAG 结构-功能关系的进展，这一研究领域正变得越来越重要。

项目成果

Molecular cloning of the heparin/heparan sulfate delta 4,5 unsaturated glycuronidase from Flavobacterium heparinum, its recombinant expression in Escherichia coli, and biochemical determination of its unique substrate specificity.

• DOI: 10.1021/bi012147o
• 发表时间: 2002-05
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: James R. Myette;Z. Shriver;T. Kiziltepe;M. W. McLean;G. Venkataraman;R. Sasisekharan

Emerging views of heparan sulfate glycosaminoglycan structure/activity relationships modulating dynamic biological functions.

硫酸乙酰肝素糖胺聚糖结构/活性关系调节动态生物功能的新观点。

• DOI: 10.1016/s1050-1738(01)00150-5
• 发表时间: 2002
• 期刊: Trends in cardiovascular medicine
• 影响因子: 9.3
• 作者: Shriver,Zachary;Liu,Dongfang;Sasisekharan,Ram
• 通讯作者: Sasisekharan,Ram

Glycan receptor binding of the influenza A virus H7N9 hemagglutinin.

• DOI: 10.1016/j.cell.2013.05.034
• 发表时间: 2013-06-20
• 期刊: Cell
• 影响因子: 64.5
• 作者: Tharakaraman K;Jayaraman A;Raman R;Viswanathan K;Stebbins NW;Johnson D;Shriver Z;Sasisekharan V;Sasisekharan R
• 通讯作者: Sasisekharan R

Biochemical characterization of the chondroitinase B active site.

软骨素酶 B 活性位点的生化特征。

• DOI: 10.1074/jbc.m201552200
• 发表时间: 2002
• 期刊: The Journal of biological chemistry
• 作者: Pojasek,Kevin;Raman,Rahul;Kiley,Patrick;Venkataraman,Ganesh;Sasisekharan,Ram
• 通讯作者: Sasisekharan,Ram

Mass spectrometric and capillary electrophoretic investigation of the enzymatic degradation of heparin-like glycosaminoglycans.

肝素样糖胺聚糖酶降解的质谱和毛细管电泳研究。

• DOI: 10.1073/pnas.95.8.4176
• 发表时间: 1998
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Rhomberg,AJ;Ernst,S;Sasisekharan,R;Biemann,K
• 通讯作者: Biemann,K