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）：在真核细胞的细胞 - 纤维细胞基质（ECM）界面中，糖胺聚糖（GAG）是复杂的多糖。在理解细胞ECM相互作用的动态性质的重要性（历史上，ECM被认为是一种使细胞水化的惰性材料）在影响细胞和高阶组织和器官水平的表型中的重要性。该范式的核心是化学异质性插科打s和众多蛋白质在细胞外环境中的特定相互作用。因此，理解插科打的结构功能关系已成为一个基本领域及其在鉴定新型治疗靶标的潜力的重要性。然而，由于插科打g的多分散性和化学异质性，过去朝着这个目标的进展受到限制，这对它们的隔离和表征构成了许多挑战。几年来，我们一直集中精力解决这些挑战，以开发酶促工具，以可预测的GAG解放以解码其序列信息以及敏感的分析技术，以准确地检测和表征来自细胞或组织可用的极少量的GAG。专注于两种重要类插科打s。 HSGAG和CSGAGS我们成功地证明了工具的实用性，以开发快速，强大的技术来测序插科打s。在这项研究中，我们试图扩展工具的开发，以关注HSGAG和CSGAG结构 - 功能关系的生化和生物学方面。最后，我们试图开发一个数据库平台，以捕获和传播与堵嘴结构功能关系有关的信息和数据，这是在插科打gag研究领域的背景下的一种新方法。我们认为，我们的研究将真正加快理解基本生物过程中插科打结构功能关系的进步 - 一个正在越来越重要的研究领域。

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

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