Structural mechanisms of fibroblast growth factor receptor signalling at the membrane interface

成纤维细胞生长因子受体(FGFRs)是一种受体酪氨酸激酶，在胚胎发育、组织重塑、伤口愈合和癌症中发挥关键作用。FGFR信令的一个持久的谜团是如何根据输入信号的性质和水平(特定成纤维细胞生长因子变体的结合)来选择输出(特定信令模块的激活，例如Raf-MEK-ERK)。在这个项目中，你将结合结构生物学和生物物理学(天然MS和结构蛋白质组学)方法以及诱变、细胞生物学和成像来阐明激酶结构域、膜旁区域和膜脂之间相互作用的关键机制特征，并确定这些相互作用如何通过结构性结合的FGFR接头蛋白FRS2影响下游信号传递。具体来说，你将：1.在膜-模拟系统(例如纳米盘)中使用核磁共振表征膜脂与FGFR3的JM-KD之间的相互作用2.使用利兹开创的天然质谱学(MS)和结构蛋白质组学方法，包括离子迁移率(IM-MS)，氢-氚交换(HDX-MS)和蛋白质的快速光化学氧化(FPOP)，以提供脂-蛋白质相互作用的正交结构图以及天然表达的FGFRs周围的脂组成的特征。阐明FGFR3 JM-KD和膜锚定适配蛋白FRS2如何使用核磁共振、MS和基于细胞的成像方法相互作用。学生将受益于阿斯特伯里中心世界领先的结构生物学基础设施，最近投资2000万英镑，专业设备包括950 MHz核磁共振、高端Orbitrap蛋白质质谱仪和泰坦Krios冷冻-EM。利兹和阿斯利康在FGFR细胞生物学和结构生物学方面拥有协同专业知识，可以为该项目提供尖端基础设施，为学生提供独特的竞争优势，具有非常独特的研究经验，可以有效地利用未来的职业机会，无论是学术还是工业。

Fibroblast growth factor receptors (FGFRs) are receptor tyrosine kinases that play key roles in embryo development, tissue remodelling, wound healing, and cancer. An enduring enigma of FGFR signalling is the question of how the output (activation of a specific signalling module, e.g. Raf-Mek-Erk) is selected based on the nature and level of the input signal (binding of a particular FGF variant). In this project, you will combine structural biology and biophysical (native MS & structural proteomics) approaches together with mutagenesis, cell biology and imaging to elucidate key mechanistic features of the interaction between the kinase domain, juxtamembrane region and membrane lipids, and determine how these interactions influence downstream signalling through the constitutively-bound FGFR adaptor protein, FRS2.Specifically, you will:1. Characterise the interaction between membrane lipids and the JM-KD of FGFR3 using NMR in a membrane-mimetic system (e.g. nanodiscs)2. Use native mass spectrometry (MS) & structural proteomics methods pioneered in Leeds, including ion-mobility (IM-MS), hydrogen-deuterium exchange (HDX-MS) and fast photochemical oxidation of proteins (FPOP), to provide orthogonal structural mapping of lipid-protein interactions as well as characterisation of the lipid composition around natively-expressed FGFRs.3. Elucidate how FGFR3 JM-KD and the membrane-anchored adapter protein FRS2 interact using NMR, MS and cell-based imaging approaches.The student will benefit from the Astbury Centre's world-leading structural biology infrastructure, with >£20M of recent investment, with specialist equipment including 950 MHz NMR, top-end Orbitrap protein mass spectrometry and Titan Krios cryo-EM.Leeds and AstraZeneca have synergistic expertise in FGFR cell biology and structural biology and together can provide cutting-edge infrastructure to the project and provide a unique competitive advantage to the student, with a highly distinctive research experience that can be effectively levered for future career opportunities, whether academic or industrial.