PROTEIN CRYSTAL STRUCTURE DETERMINATIONS RELATED TO NITRIC OXIDE SIGNALING

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 我们正在申请束流时间，以进一步推进四个PI实验室中正在进行的项目。这些项目包括(1)参与一氧化氮信号转导的蛋白质的结构研究，重点是可溶性鸟苷环化酶(SGC)、其功能结构域及其调控伙伴。已经表达了几个sGC片段，并获得了其中两个的小晶体。两个模型系统，亚硝酚4(Rhodnius Prolixus)和硫氧还蛋白(人)正在进行研究，以了解亚硝酰血红素和S亚硝基半胱氨酸的化学生物学。这些蛋白质的晶体绕射超过1。(2)参与铜运输和稳态的蛋白质的结构(McEway)；(3)Cro和Lipocalin蛋白质的进化(Cordes)；以及(4)Qed，(Bandarian)晶体的结构/机制研究可供所有项目使用。同步辐射用于(1)MAD结构测定，(2)提供足够的分辨率来确定精确的几何参数，如血红素扭曲、金属结合位几何和S-亚硫醇几何构型，(3)提供可变波长的X射线用于明确识别结合金属原子和(4)控制金属中心和S-NO键的光还原。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We are requesting beam time to further on-¿going projects in the laboratories of four PIs. These projects include (1) Structural studies of proteins involved in nitric oxide signaling, with emphasis on soluble guanylate cyclase (sGC), its functional domains and its regulatory partners. Several sGC fragments have been expressed and small crystals obtained for two of these. Two model systems, nitrophorin 4(Rhodnius prolixus) and thioredoxin (human)are under study for understanding nitrosyl heme and S-¿nitrosocysteine chemical biology. Crystals of these proteins diffract past 1.¿.(2)Structures of proteins involved in copper transport and homeostasis(McEvoy);(3) Evolution of Cro and lipocalin proteins (Cordes), and (4) Structure/mechanism studies of QueD,(Bandarian)Crystals are available for all projects. Synchrotron radiation is needed for(1)MAD structure determination, (2) to provide sufficient resolution to determine accurate geometrical parameters such as heme distortion, metal binding site geometry and S-nitrosothiol geometry;(3) to provide variable wavelength x-¿rays for unambiguous identification of bound metal atoms and (4)to control photoreduction of metal sites and S-¿NO bonds.