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）：该项目重点研究人类透明质酸酶-1 (hHyal-1) 和透明质酸酶-2 (hHyal-2) 在介导细胞外基质多糖透明质酸 (HA) 所承担的生物功能中的作用。 HA 充当将细胞粘合在一起的“粘合剂”。为了使细胞移入或移出基质，HA 必须被 hHyals 打碎。受损组织的修复需要细胞在基质内移动，异常细胞的逃逸（转移）也是如此。 HA 片段被邻近细胞检测到，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 Hyals 产生的 HA 片段的大小。 hHyal-1 和 hHyal-2 在组织炎症和癌症中发挥着关键作用，因此具有特殊的医学重要性，因此是开发新型抗癌和抗炎疗法的绝佳靶点。直到最近，由于与活性 N-糖基化 hHyals 的高产率生产相关的技术挑战，这些酶的结构功能研究还不可能实现。 Herzberg 实验室最近开发了一种 hHyal-1 和 hHyal-2 的制备方法，并确定了 hHyal-1 的晶体结构。现在该阶段将采用体外方法来准确定义 hHyal-1 和 hHyal-2 的催化功能和抑制作用，以及它们与蛋白质伙伴、CD44（透明质酸结合蛋白）、受 hHyal-2 调节的 RON 受体酪氨酸激酶以及激活 RON 并使用 hHyal-2 附着到宿主细胞的 Jaagsiekte 绵羊逆转录病毒包膜蛋白的相互作用。该提案中提出的研究计划将在原子水平上阐明透明质酸酶控制的复杂细胞过程背后的分子机制，并将为新疗法的开发提供结构基础。公共健康相关性：透明质酸酶 1 和透明质酸酶 2 通过其控制细胞外多糖透明质酸 (HA) 周转的酶活性，对细胞外基质的完整性和功能至关重要。 HA 周转处于微妙的平衡状态，透明质酸酶活性升高会导致炎症性疾病以及癌症进展和侵袭。该项目旨在在体外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，并为开发透明质酸酶作为抗癌和抗炎药物靶点奠定基础。