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）界面处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被认为是使细胞水合的惰性材料）在细胞和更高级组织和器官水平上影响表型的重要性方面已经有了范式转变。这种模式的核心是化学异质性GAG与细胞外环境中的许多蛋白质之间的特异性相互作用。因此，了解糖胺聚糖的结构-功能关系作为一个基本领域及其在鉴定新的治疗靶点方面的潜力已经变得重要。然而，在过去，由于GAG的多分散性和化学异质性对其分离和表征提出了许多挑战，因此实现这一目标的进展有限。几年来，我们一直致力于解决这些挑战，开发用于GAG可预测解聚的酶促工具，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。聚焦于两个重要类别的GAG，即HSGAG和CSGAG，我们已经成功地证明了我们的工具在开发用于测序GAG的快速和稳健的技术方面的实用性。在这项研究中，我们寻求扩大我们的工具的发展，专注于HSGAG和CSGAG结构-功能关系的生物化学和生物学方面。最后，我们寻求开发一个数据库平台，以捕获和传播有关GAG结构功能关系的信息和数据，这是一种新的方法，在GAG研究领域的背景下。我们相信，我们的研究将真正加快了解GAG的基本生物过程中的结构功能关系的进展，这是一个研究领域，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

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

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

The soft palate is an important site of adaptation for transmissible influenza viruses.

• DOI: 10.1038/nature15379
• 发表时间: 2015-10-01
• 期刊: Nature
• 影响因子: 64.8
• 作者: Lakdawala SS;Jayaraman A;Halpin RA;Lamirande EW;Shih AR;Stockwell TB;Lin X;Simenauer A;Hanson CT;Vogel L;Paskel M;Minai M;Moore I;Orandle M;Das SR;Wentworth DE;Sasisekharan R;Subbarao K
• 通讯作者: Subbarao K

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