PROBING THE ROLES OF CALCIUM AND SULFUR IN PROSTAGLANDIN SYNTHASE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 前列腺素(PG)D2作为一种主要的前列腺素类物质存在于多种组织中。在中枢神经系统中，PGD2调节机体的各种反应，如诱导睡眠、体温、激素释放等。在前列腺素D2合成(PGDS)的两种类型中，造血型PGDS(H-PGDS)定位于多种组织的Th2细胞和肥大细胞中，这需要谷胱甘肽(GSH)来催化反应。最近有报道称，GSH结合部位(催化口袋)被钙离子激活。我们将比较存在和不存在GSH的情况下的CaXAS。在这个建议中，我们研究了钙在酶反应中的结构和功能作用。为了探索功能钙结合部位，我们将对每二聚体1个钙的H-PGDS样品进行CaXAS研究，并从大肠杆菌中表达和纯化H-PGDS样品。还将探讨其他二价金属和过渡金属可能的功能作用。此外，我们计划进行SAXS来研究GSH在结合口袋中的结合形式。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Prostaglandin (PG) D2 is produced in a variety of tissues as a major prostanoid. In the central nervous system, PGD2 is known to regulate various body reaction, such as inducing sleep, body temperature, hormone release. Among the two types of Prostaglandin D2 synthese (PGDS), hematopoietic PGDS (H-PGDS) is localized in Th2 cells and mast cells in a variety of tissues, which requires glutathione (GSH) for the catalytic reaction. It is reported recently that the GSH binding site (catalytic pocket) is activated by the Ca2+ ions. We will compare Ca XAS in the presence and absence of GSH. In this proposal, we study the structural and functional roles of Ca in the enzymatic reaction. To probe the functional Ca binding site, we will perform Ca XAS studies on H-PGDS samples having 1 Ca per dimer H-PGDS samples expressed and purified from E. coli. Possible functional roles of other divalent metals and transition metals will also be explored. Additionally, we plan to carry out SAXS to study the bonding form of GSH in the binding pocket.