Structure-Function studies of human hyaluronidases

人类透明质酸酶的结构-功能研究

• 批准号: 7778818
• 负责人: OSNAT HERZBERG
• 金额: $ 31.94万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7778818
• 关键词: Abnormal Cell; Active Sites; Affinity; Anti-Inflammatory Agents; Anti-inflammatory; Binding; Biochemical; Biological Process; CD44 Antigens; CD44 gene; Calorimetry; Catalysis; Cell physiology; Cells; Characteristics; Complex; Data; Degenerative polyarthritis; Development; Disease; Drug Delivery Systems; Embryonic Development; Enzymes; Equilibrium; Exhibits; Experimental Designs; Extracellular Domain; Extracellular Matrix; Foundations; Glues; Goals; Human; Hyaluronan; Hyaluronidase; In Vitro; Inflammation; Inflammatory; Kinetics; Light; Link; Malignant Neoplasms; Mediation; Medical; Methods; Molecular; Neoplasm Metastasis; Ovine pulmonary adenocarcinoma virus; Physiological; Play; Polymers; Polysaccharides; Preparation; Process; Production; Proteins; Receptor Protein-Tyrosine Kinases; Recombinants; Research; Rheumatoid Arthritis; Roentgen Rays; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Specificity; Staging; Structure; Surface; Surface Plasmon Resonance; Therapeutic; Therapeutic Agents; Thermodynamics; Tissues; Titrations; Wound Healing; base; cell motility; design; env Gene Products; extracellular; human MST1R protein; human disease; in vivo; inhibitor/antagonist; injured; novel; novel therapeutics; overexpression; protein structure; public health relevance; receptor; repaired; response; tumor progression

DESCRIPTION (provided by applicant): This project focuses on the roles of human hyaluronidase-1 (hHyal-1) and hyaluronidase-2 (hHyal-2) in the mediation of the biological functions assumed by the extracellular matrix polysaccharide hyaluronan (HA). HA serves as the "glue" that binds cells together. In order for cells to move in or out of the matrix, HA must be fragmented by hHyals. Repair of injured tissue requires cell movement within the matrix, as does the escape of an abnormal cell (metastasis). HA fragments are detected by neighboring cells, and an intracellular response occurs via signal transduction pathways. The signal received is dependent on the size of the HA fragments as produced by the Hyals. Of particular medical importance, hHyal-1 and hHyal-2 play key roles in tissue inflammation and in cancer, and therefore are excellent targets for the development of novel anti-cancer and anti-inflammatory therapeutics. Until recently, structure-function studies of these enzymes have not been possible owing to the technical challenge associated with high yield production of the active N-glycosylated hHyals. A method for hHyal-1 and hHyal-2 preparation has been recently developed in the Herzberg lab and the crystal structure of hHyal-1 has been determined. The stage is now set to employ in vitro methods to accurately define hHyal-1 and hHyal-2 catalytic function and inhibition, and their interactions with protein partners, CD44 - the hyaluronan biding protein, RON receptor tyrosine kinase that is regulated by hHyal-2, and Jaagsiekte sheep retrovirus envelope protein that activates RON and uses hHyal-2 to attach to the host cell. The research plan set forth in this proposal will illuminate at the atomic level the molecular mechanisms underlying the complex cellular processes controlled by the hyaluronidases and will provide the structural foundation for the development of new therapeutics. PUBLIC HEALTH RELEVANCE: Hyaluronidase-1 and hyaluronidase-2 are crucial to the integrity and functioning of the extracellular matrix through their enzymatic activity that controls the turnover of the extracellular polysaccharide hyaluronan (HA). The HA turnover is delicately balanced and heightened hyaluronidase activity leads to inflammatory diseases and cancer progression and invasion. This project seeks to characterize the human hyaluronidases and their interactions with cellular partner proteins in vitro to shed light on their in vivo function and to lay the foundation for development of the hyaluronidases as anti cancer and anti inflammatory drug targets.

描述（由申请人提供）：该项目着重于人透明质酸酶-1（HHYY-1）和透明质酸酶-2（HHYY-2）在介导的生物学功能中的作用。 HA充当将细胞结合在一起的“胶”。为了使细胞进出矩阵，HA必须被Hhyals碎裂。受伤组织的修复需要在基质内运动，异常细胞的逃生（转移）也是如此。邻近细胞检测到HA片段，并通过信号转导途径发生细胞内反应。接收到的信号取决于透明产生的HA片段的大小。尤其重要的是，HHYAL-1和HHYY-2在组织炎症和癌症中起关键作用，因此是开发新型抗癌和抗炎治疗剂的绝佳靶标。直到最近，由于与活性N-糖基化的Hhyals的高产量产生有关的技术挑战，这些酶的结构功能研究还没有可能。最近在HERZBERG LAB中开发了HHYY-1和HHYY-2制备方法，并确定了HHYY-1的晶体结构。 The stage is now set to employ in vitro methods to accurately define hHyal-1 a​​nd hHyal-2 catalytic function and inhibition, and their interactions with protein partners, CD44 - the hyaluronan biding protein, RON receptor tyrosine kinase that is regulated by hHyal-2, and Jaagsiekte sheep retrovirus envelope protein that activates RON and uses hHyal-2 to attach to the host cell.该提案中规定的研究计划将在原子水平上阐明由透明质酸酶控制的复杂细胞过程的分子机制，并将为开发新疗法的结构基础提供结构基础。公共卫生相关性：透明质酸酶-1和透明质酸酶-2通过其酶促活性控制细胞外基质的完整性和功能至关重要，该酶活性控制细胞外多糖透明质酸（HA）的周转。 HA的周转率是微妙的平衡，透明质酸酶活性提高导致炎症性疾病以及癌症的进展和侵袭。该项目旨在表征人类透明质酸酶及其与细胞伴侣蛋白的相互作用，以阐明其体内功能，并为透明质酸酶作为抗癌和抗炎性药物靶标的发展奠定基础。