Functional phosphosignaling in Mtb infection

结核分枝杆菌感染中的功能性磷酸信号传导

• 批准号: 10177868
• 负责人: Christoph Grundner
• 金额: $ 21.15万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-02 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10177868
• 关键词: Affect; Biological; Cells; Complement; Data; Data Set; Docking; Endopeptidase K; Enzyme-Linked Immunosorbent Assay; Equipment and supply inventories; Event; Foundations; Global Change; Human; Immune; Immune response; Immunity; Individual; Infection; Knock-out; Lead; Link; Lip structure; Maps; Mass Spectrum Analysis; Measurement; Measures; Mediating; Methods; Modeling; Molecular Conformation; Mutation; Mycobacterium tuberculosis; Natural Immunity; Outcome; Pathway interactions; Pattern; Peptide Hydrolases; Peptides; Phenotype; Phosphoproteins; Phosphorylation; Phosphorylation Site; Phosphotransferases; Play; Predisposition; Primary Infection; Proteins; Proteolysis; Proteome; Proteomics; RNA Interference; Role; Sampling; Shotguns; Signal Transduction; Site; Structure; Surface; System; Testing; Trypsin; Tuberculosis; base; cytokine; insight; knock-down; macrophage; mimetics; mutant; phosphoproteomics; protein protein interaction; protein structure; protein structure function; response; targeted treatment; three dimensional structure; tool

PROJECT SUMMARY Phosphosignaling coordinates much of the host macrophage’s (M’s) response to infection with Mycobacterium tuberculosis (Mtb). Surprisingly, only few studies to date have measured global changes in the phosphoproteome upon infection with Mtb by mass spectrometry, and none in primary human Ms. In addition, a major challenge in the cell signaling field today is to separate phosphorylation events that are functional from those that are not. The lack of tools, experimental or computational, to d istinguish functional from non-functional phosphorylation events is the reason why many current studies of phosphosignaling largely remain inventories of phosphosites that provide limited biological insight. Combining phospho- and structural proteomics, we will test the hypothesis that the occurrence of a structural change of a protein upon phosphorylation is a reliable predictor of functional phosphorylation events. We will test this idea by combining detailed phosphosite mapping and global protein structure measurements to compare the proteomes of uninfected to Mtb-infected primary human Ms. We will correlate changes in individual phosphorylation sites with changes in the proteolytic cleavage pattern as a readout of structural change. Phosphorylation events that change a protein’s structure will be tested for their function in Mtb infection. We will knock out (or down) the phosphoprotein and test the effect on bacterial loads and M function. To link effects of knockout to specific phosphorylation sites, we will complement with the protein bearing phosphoablative and -mimetic mutations and test for effects on M signaling and Mtb infection. This project will provide a proof-of-principle for an experimental method to identify functional phosphorylation sites in general, and in particular in the context of the Mtb-infected M. These data will allow for more meaningful curation of M phosphorylation datasets, provide a global, functional view of the cell’s remodeling upon infection, highlight the specific immune pathways triggered by Mtb infection, identify potential new host-directed therapy targets, and identify downstream effectors that control or exacerbate Mtb infection.

项目摘要 磷酸化信号传导协调宿主巨噬细胞（M β）对感染的大部分反应， 结核分枝杆菌（Mtb）。令人惊讶的是，迄今为止，只有少数研究测量了全球变化， 通过质谱分析，在感染Mtb后的磷酸化蛋白质组中，没有磷酸化蛋白质组，而在原代人Mtb中没有。在 此外，当今细胞信号传导领域的一个主要挑战是分离磷酸化事件， 功能性的区别。缺乏工具，实验或计算，以区分功能 这就是为什么许多目前关于磷酸化信号的研究 大部分仍然是提供有限生物学见解的磷酸盐的库存。结合磷酸和 结构蛋白质组学，我们将测试的假设，发生结构变化的蛋白质后， 磷酸化是功能性磷酸化事件的可靠预测因素。我们将通过结合 详细的磷酸酶定位和全球蛋白质结构测量，以比较蛋白质组的 未感染的人结核分枝杆菌与感染结核分枝杆菌的原代人结核分枝杆菌。我们将把单个磷酸化位点的变化 蛋白水解裂解模式的变化作为结构变化的读数。磷酸化事件， 改变蛋白质的结构将测试它们在结核分枝杆菌感染中的功能。我们将在这里， 磷蛋白，并测试对细菌负荷和M β功能的影响。将基因敲除的效果与 磷酸化位点，我们将与带有磷酸化和拟磷酸化突变的蛋白互补。 并测试对Mtb信号传导和Mtb感染的影响。该项目将提供一个原理证明， 实验方法，以确定功能磷酸化位点一般，特别是在上下文中， Mtb感染的M.这些数据将允许对M β磷酸化数据集进行更有意义的管理， 提供感染后细胞重塑的整体功能视图，突出特定的免疫途径 确定潜在的新的宿主定向治疗靶点，并确定下游靶点。 控制或加重Mtb感染的效应物。

项目成果

Dual Inhibition of Mycobacterium tuberculosis and the Host TGFBR1 by an Anilinoquinazoline.

Anilinoquinazoline 对结核分枝杆菌和宿主 TGFBR1 的双重抑制。

• DOI: 10.1021/acs.jmedchem.3c01273
• 发表时间: 2023
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Nandakumar,Meganathan;Ollodart,Anja;Fleck,Neil;Kapadia,NiravR;Frando,Andrew;Boradia,Vishant;Smith,JefferyL;Chen,Junxi;Zuercher,WilliamJ;Willson,TimothyM;Grundner,Christoph
• 通讯作者: Grundner,Christoph