PROBING THE ROLES OF CALCIUM AND SULFUR IN PROSTAGLANDIN SYNTHASE

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 前列腺素（PG）D2作为一种主要的前列腺素在多种组织中产生。在中枢神经系统中，PGD 2被认为可以调节各种身体反应，如诱导睡眠、体温、激素释放。在前列腺素D2合成（PGDS）的两种类型中，造血PGDS（H-PGDS）定位于多种组织中的Th 2细胞和肥大细胞，其需要谷胱甘肽（GSH）进行催化反应。最近有报道称，GSH结合位点（催化口袋）被Ca ~（2+）离子激活。我们将比较存在和不存在GSH的情况下的Ca XAS。在这个提议中，我们研究了钙在酶促反应中的结构和功能作用。为了探测功能性Ca结合位点，我们将对从大肠杆菌表达和纯化的每个二聚体具有1个Ca的H-PGDS样品进行Ca XAS研究。杆菌其他二价金属和过渡金属可能的功能作用也将探讨。此外，我们计划进行小角X射线散射来研究谷胱甘肽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.