PROTEIN CRYSTAL STRUCTURE DETERMINATIONS RELATED TO NITRIC OXIDE SIGNALING

与一氧化氮信号传导相关的蛋白质晶体结构测定

• 批准号: 8362426
• 负责人: SUE L ROBERTS
• 金额: $ 0.03万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362426
• 关键词: Binding; Biological Models; Biology; Chemicals; Copper; Evolution; Funding; Grant; Heme; Homeostasis; Laboratories; Metal Binding Site; Metals; National Center for Research Resources; Nitric Oxide; Principal Investigator; Proteins; Radiation; Research; Research Infrastructure; Resolution; Resources; Rhodnius; S-Nitrosothiols; Signal Transduction; Site; Soluble Guanylate Cyclase; Source; Structure; Time; United States National Institutes of Health; cost; human TXN protein; nitrophorin; protein structure; structural biology; synchrotron radiation

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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们正在请求束流时间，以进一步在四个PI的实验室中进行的项目。这些项目包括（1）参与一氧化氮信号传导的蛋白质的结构研究，重点是可溶性鸟苷酸环化酶（sGC），其功能结构域及其调控伙伴。已经表达了几个sGC片段，并获得了其中两个片段的小晶体。两个模型系统，nitrophorin 4（Rhodnius prolixus）和硫氧还蛋白（人类）正在研究中，以了解亚硝基血红素和S-亚硝基半胱氨酸的化学生物学。这些蛋白质的晶体会在1分钟后发生作用。（2）参与铜转运和体内平衡的蛋白质结构（McEhrs）;（3）Cro和脂质运载蛋白的进化（Cordes）;（4）QueD的结构/机制研究，（Bandarian）晶体可用于所有项目。同步辐射需要用于（1）MAD结构测定，（2）提供足够的分辨率以确定准确的几何参数，如血红素畸变，金属结合位点几何形状和S-亚硝基硫醇几何形状;（3）提供可变波长的X-射线以明确识别结合的金属原子和（4）控制金属位点和S-NO键的光还原。