X-RAY DIFFRACTION ANALYSIS OF METABOLITE SENSING RIBOSWITCHES, G-PROTEIN/SMALL M

代谢物传感核开关、G 蛋白/小 M 的 X 射线衍射分析

• 批准号: 8170271
• 负责人: Joseph E Wedekind
• 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/8170271
• 关键词: Affect; Affinity; Binding; Biology; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Funding; GTP-Binding Proteins; Goals; Grant; HIV-1; Human; Institution; Learning; Ligase; Molecular; Proteins; Public Health; Regulator Genes; Research; Research Personnel; Resolution; Resources; Signal Transduction; Source; Structure; Techniques; United States National Institutes of Health; Viral; Work; X ray diffraction analysis; X-Ray Diffraction; antimicrobial; novel; phosducin; small molecule; vif Gene Products

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. The goal of this proposal is to collect high-resolution X-ray diffraction data for three different biomacromolecules that are relevant to biology and public health. Our first objective will be to learn the mode of metabolite recognition by a bacterial, PreQ1 riboswitch. Such information is essential to understand this novel gene regulatory paradigm, which may serve as an antimicrobial target. Previously we determined the structure of the PreQ1 riboswitch at SSRL and have now obtained new crystals grown in the presence of various metabolites that span a broad range of binding affinities (Kd = 5 nM to 500 nM). We are also investigating the mode of binding of the G-protein beta-gamma to various small molecules shown to affect cell signaling by our collaborator. These molecules are hypothesized to bind in a "hotspot" on the beta subunit. Our goal is to locate this molecules by difference Fourier techniques, which has implications for developing novel effectors of G-protein signaling. Finally, we are working to determine the crystal structure of the HIV-1 protein Vif in complex with components of the Cullin-RING ligase complex. This viral/host protein interaction has not been characterized at the molecular level. Our goal is to identify well-diffracting crystals in order to proceed with a structure determination. Our complex comprises two human proteins, EloB and EloC, as well as HIV-1 Vif.

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。子项目及 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。 该提案的目标是收集与生物学和公共卫生相关的三种不同生物大分子的高分辨率 X 射线衍射数据。我们的首要目标是了解细菌 PreQ1 核糖开关识别代谢物的模式。这些信息对于理解这种新的基因调控范式至关重要，它可以作为抗菌靶点。之前我们在 SSRL 确定了 PreQ1 核糖开关的结构，现在已经获得了在各种代谢物存在下生长的新晶体，这些代谢物具有广泛的结合亲和力（Kd = 5 nM 至 500 nM）。我们还在研究 G 蛋白 β-γ 与各种小分子的结合模式，我们的合作者已证明这些小分子会影响细胞信号传导。 假设这些分子结合在 β 亚基上的“热点”上。 我们的目标是通过差异傅里叶技术来定位这种分子，这对于开发新型 G 蛋白信号传导效应器具有重要意义。 最后，我们正在努力确定 HIV-1 蛋白 Vif 与 Cullin-RING 连接酶复合物成分复合物的晶体结构。这种病毒/宿主蛋白相互作用尚未在分子水平上得到表征。我们的目标是识别衍射良好的晶体，以便进行结构测定。 我们的复合物包含两种人类蛋白 EloB 和 EloC，以及 HIV-1 Vif。