CONFORMATIONAL CHANGES OF SOLUBLE QUANYLATE CYCLES

可溶性季铵盐环的构象变化

• 批准号: 8170220
• 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: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170220
• 关键词: Binding; Cells; Chemistry; Computer Retrieval of Information on Scientific Projects Database; DNA Sequence Rearrangement; Funding; Grant; Heme; Insecta; Institution; Length; Ligand Binding; Manduca sexta; N-terminal; Proteins; Research; Research Personnel; Resolution; Resources; Shapes; Signal Transduction; Soluble Guanylate Cyclase; Source; Structure; United States National Institutes of Health; analytical ultracentrifugation; cGMP production; receptor

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. Soluble guanylate cyclase (sGC), a 150 kD heterodimeric protein, is the primary NO receptor in the cell. NO binds to heme in the N-terminal domain of the ? subunit, stimulates the production of cGMP and leads to NO-dependent signaling cascades. Structure determination has been hindered by our inability to crystallize the protein. We can prepare, in high yield, a truncated sGC (msGC-NT2, 97 kD) from the insect Manduca sexta that lacks the cyclase domain. The NO chemistry of this construct is identical to that of the full-length protein. (Hu, et.al., J. Biol. Chem., (2008), 20968.). Analytical ultracentrifugation (AUC) shows that msGC-NT2 changes shape when ligands bind. SAXS studies will further define the low resolution structure of our sGC construct and determine whether significant domain rearrangement occurs when ligands bind.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 可溶性鸟苷酸环化酶 (sGC) 是一种 150 kD 异二聚体蛋白，是细胞中的主要 NO 受体。 NO 与 ? N 端结构域中的血红素结合。亚基，刺激 cGMP 的产生并导致 NO 依赖性信号级联反应。 由于我们无法使蛋白质结晶，结构测定受到阻碍。 我们可以从缺乏环化酶结构域的昆虫 Manduca sexta 中高产地制备截短的 sGC（msGC-NT2，97 kD）。 该构建体的 NO 化学成分与全长蛋白质的 NO 化学成分相同。 (Hu, et.al., J. Biol. Chem., (2008), 20968.)。分析超速离心 (AUC) 显示配体结合时 msGC-NT2 会改变形状。 SAXS 研究将进一步定义我们的 sGC 构建体的低分辨率结构，并确定配体结合时是否会发生显着的结构域重排。