Structure and functional mechanisms of molecular chaperones and protein kinases

分子伴侣和蛋白激酶的结构和功能机制

• 批准号: 9920188
• 负责人: CHARALAMPOS KALODIMOS
• 金额: $ 75.38万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9920188
• 关键词: Actins; Address; Affinity; Architecture; BCR/ABL fusion gene; Biochemical; Biological Process; Cell Adhesion; Cells; Chimeric Proteins; Chronic Myeloid Leukemia; Client; Complex; DNA Damage; Drug resistance; Human Genome; Mediating; Molecular Chaperones; Molecular Structure; NMR Spectroscopy; Pharmaceutical Preparations; Phosphotransferases; Protein Dynamics; Protein Family; Protein Kinase; Protein Tyrosine Kinase; Proteins; Proteome; Reporting; Resolution; Role; Structure; Therapeutic Intervention; Work; biophysical techniques; cell motility; fascinate; gene product; non-Native; novel; pathogenic bacteria; protein aggregation; resistance mutation; response; targeted treatment

PROJECT SUMMARY Our lab works on two main directions: first, the determination of the structural and dynamic basis for the function and assembly of large protein machineries; and second, the determination of the role of internal protein dynamics in regulating protein activity and allosteric interactions. We propose to use NMR spectroscopy, together with other biochemical and biophysical techniques, to determine at the atomic resolution the mechanisms underpinning the function of two important protein families: molecular chaperones and protein kinases. Molecular chaperones are central to maintaining a functional proteome in the cell by rescuing non- native proteins from aggregation and misfolding and assisting with their folding. Our lab reported the first ever high resolution structures of molecular chaperones in complex with unfolded proteins. We will determine the structures of important chaperones such as the Hsp40, Hsp70 and Hsp90 in complex with client proteins. We wish to address how different chaperones engage non-native proteins and how distinct chaperone architectures may alter activity. The Abl kinase holds a prominent place among the over 500 protein kinases encoded by the human genome. Abl mediates its function by participating in a number of biological processes, including actin remodeling, cell adhesion and motility, DNA damage response, and bacterial pathogen response. The Bcr-Abl fusion gene product has constitutive tyrosine kinase activity and causes chronic myeloid leukemia (CML). We will use NMR to provide fascinating, novel information about the regulatory and activation mechanisms of this important kinase. We will study how drug-resistance mutations exert their effect in lowering the drug affinity for Abl.

项目总结 我们的实验室主要在两个方向上工作：first，确定结构和动力基础 大型蛋白质机器的功能和组装；第二，确定蛋白质机器的作用 调节蛋白质活性和变构相互作用的蛋白质内部动力学。我们建议使用 核磁共振光谱学，以及其他生化和生物物理技术，以确定在 原子分辨率支持两个重要蛋白质家族功能的机制： 分子伴侣和蛋白激酶。 分子伴侣是维持细胞中功能蛋白质组的核心，它通过拯救非 天然蛋白质的聚集和错误折叠，并协助它们的折叠。我们的实验室报告说 fi首次在与未折叠蛋白质的络合物中实现分子伴侣的高分辨结构。我们 将确定重要的伴侣，如Hsp40，Hsp70和Hsp90的结构 与客户蛋白形成复合体。我们希望解决不同的监护人如何与非本地人接触 蛋白质，以及不同的伴侣结构如何改变活性。 Abl激酶在人类编码的500多种蛋白激酶中占有重要地位。 基因组。ABL通过参与许多生物过程来调节其功能，包括 肌动蛋白重塑、细胞黏附和运动、DNA损伤反应和细菌病原体 回应。Bcr-abl融合基因产物具有结构性酪氨酸激酶活性，可引起慢性 髓系白血病(CML)。我们将使用核磁共振来提供有关 这一重要的激酶的调节和激活机制。我们将研究抗药性是如何 突变在降低ABL的药物fi含量方面发挥了作用。