The chaperone cycle of fibroblast growth factor receptor kinases in molecular detail

成纤维细胞生长因子受体激酶的分子伴侣循环的分子细节

• 批准号: BB/W008017/1
• 负责人: Alexander Breeze
• 金额: $ 78.76万
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW008017%2F1
• 关键词: chaperone; cycle; fibroblast; growth; factor

As cells in our body age or encounter external stresses, such as toxins derived from diet or the environment, proteins within the cell can accumulate damage (for example mutations) or alterations in expression levels that can cause them to form aggregates that are damaging to cell function and survival. Diseases of aging such as Alzheimer's and Parkinson's are examples of conditions associated with the formation of large, aggregated protein structures within certain groups of neuronal cells, leading to manifestations including dementia, tremor or loss of motor control. In response to stresses, cells marshal a set of proteins called chaperones whose function is to rescue such aberrantly folded or aggregated proteins. Often, however, the chaperone response itself can become compromised through accumulation of cellular damage, which can blunt the organism's ability to remediate protein aggregation and lead to disease progression with advancing age.One such arm of the chaperone response involves the protein HSP90, which is assisted in its recognition of 'client' proteins by various co-chaperones that include Cdc37. Cdc37 is the specific co-chaperone for an important family of proteins involved in cellular signalling known as kinases. Receptor tyrosine kinases (RTKs) are a type of 'signalling' kinase that transduce signals from outside to inside cells and include fibroblast growth factor receptor kinases (FGFRs), the subject of this study. FGFRs are important in embryonic development, wound-healing, generation of new blood vessels, and in altered forms are responsible for driving several types of cancer. We are interested in establishing the molecular details of how FGFRs interact with the Cdc37-HSP90 chaperone system, and in particular in how Cdc37 is able to distinguish between forms of FGFRs that require chaperone intervention (for example, altered forms of FGFRs responsible for diseases such as cancer) and those that do not (for example, wild-type variants of most FGFRs).Our project seeks to fill in some of the important gaps in our knowledge, by using a range of cutting-edge technologies and approaches. In particular, cryo-electron microscopy (cryo-EM) has recently undergone a dramatic improvement in the level of structural detail it can provide, thanks to new technologies and methods, and in Leeds and UCL we have access to the latest-generation cryo-EM equipment and expertise. Other structural techniques such as NMR and mass spectrometry have also seen significant advances in capability, and again, we have expertise and state-of-the-art equipment in Leeds and UCL to undertake this work. Moreover, over the last 5 years we have built up a strong body of pilot data, generating the components required, in-particular the protein complexes, which are often the limiting step in these challenging projects.Understanding the molecular-level 'rules of engagement' of Cdc37 and the chaperone system with FGFR kinases and their altered forms will help us to understand not only the normal functioning of the chaperone system in respect of this important class of protein clients (RTKs), but also how it might become overwhelmed or subverted in acute or cumulative cellular stress. Such understanding can also assist in efforts to target the chaperone system selectively to treat cancers driven by oncogenic kinases that are 'addicted' to chaperone intervention to maintain their activity.

随着我们身体中的细胞老化或遇到外部压力，例如来自饮食或环境的毒素，细胞内的蛋白质会积累损伤（例如突变）或表达水平的改变，导致它们形成损害细胞功能和存活的聚集体。诸如阿尔茨海默氏症和帕金森氏症的衰老疾病是与在某些神经元细胞群内形成大的、聚集的蛋白质结构相关的病症的实例，导致包括痴呆、震颤或运动控制丧失的表现。为了应对压力，细胞排列了一组称为分子伴侣的蛋白质，其功能是拯救这些异常折叠或聚集的蛋白质。然而，通常情况下，分子伴侣反应本身会因为细胞损伤的累积而受损，这会削弱机体修复蛋白质聚集的能力，并导致疾病随着年龄的增长而进展。分子伴侣反应的一个这样的分支涉及蛋白质HSP 90，它通过包括Cdc 37在内的各种辅助分子伴侣来辅助其识别“客户”蛋白质。cdc 37是参与细胞信号传导的重要蛋白质家族（称为激酶）的特异性辅助分子伴侣。受体酪氨酸激酶（RTK）是一种“信号传导”激酶，将信号从细胞外传递到细胞内，包括本研究的主题成纤维细胞生长因子受体激酶（FGFR）。FGFR在胚胎发育、伤口愈合、新血管生成中很重要，并且以改变的形式负责驱动几种类型的癌症。我们感兴趣的是建立FGFR如何与Cdc 37-HSP 90分子伴侣系统相互作用的分子细节，特别是Cdc 37如何能够区分需要分子伴侣干预的FGFR形式（例如，改变形式的FGFR负责疾病，如癌症）和那些不我们的项目旨在通过使用一系列尖端技术和方法来填补我们知识中的一些重要空白。特别是，冷冻电子显微镜（cryo-EM）最近经历了一个戏剧性的改善，它可以提供的结构细节的水平，由于新的技术和方法，并在利兹和伦敦大学学院，我们有机会获得最新一代的cryo-EM设备和专业知识。其他结构技术，如核磁共振和质谱分析，在能力上也取得了重大进展，同样，我们在利兹和伦敦大学学院拥有专业知识和最先进的设备来开展这项工作。此外，在过去的5年里，我们已经建立了一个强大的试点数据，产生所需的组件，特别是蛋白质复合物，这通常是这些具有挑战性的项目中的限制步骤。了解分子-Cdc 37和具有FGFR激酶的分子伴侣系统及其改变形式的水平“参与规则”将帮助我们不仅理解分子伴侣系统的正常功能，这类重要的蛋白质客户端（RTK）的尊重，但也如何可能成为不堪重负或在急性或累积的细胞压力颠覆。这样的理解也可以有助于选择性地靶向分子伴侣系统以治疗由致癌激酶驱动的癌症的努力，所述致癌激酶对分子伴侣干预“上瘾”以维持其活性。