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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 150 kD的异二聚体蛋白是可溶性鸟苷酸环化酶（SGC）是细胞中的主要NO受体。在n末端结构域中没有与血红素的结合？亚基刺激CGMP的产生，并导致无依赖性信号级联。 我们无法结晶蛋白质来阻碍结构的确定。 我们可以从缺乏循环酶结构域的昆虫曼达卡·萨克斯塔（Manduca Sexta）中以高收益的高收率制备截短的SGC（MSGC-NT2，97 kd）。 该构建体的无化学性质与全长蛋白的化学性质相同。 （Hu等，Al.，J。Biol。Chem。，（2008），20968。）。分析超速离心（AUC）表明，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，并确定当配体结合时是否发生重要的结构域重排。