Advanced fluorescence spectroscopy to unravel functional oligomerization of a newly discovered Ca2+ channel and its role in heart failure.

先进的荧光光谱揭示了新发现的 Ca2 通道的功能性寡聚及其在心力衰竭中的作用。

• 批准号: 2885854
• 金额: --
• 依托单位: University of St Andrews
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2885854
• 关键词: Advanced; fluorescence; spectroscopy; unravel; functional

Endoplasmic reticulum (ER) homeostasis is becoming an increasingly interesting target in heart failure, since accumulation of misfolded or unfolded proteins can easily lead to ER stress, which in turn can play an important role in the development of cardiovascular diseases. ER transmembrane proteins can play prominent signalling roles that influence cell death or survival. Mitsugumin 23 (MG23), is a newly discovered endoplasmic/sarcoplasmic reticulum (ER/SR) transmembrane protein thought to be oligomeric in nature. We suggest that the dynamic oligomerization behaviour of MG23 controls its functional role. Our latest data suggest a role for MG23 in cardiac fibrosis, SR Ca2+ leak and the early stages of heart failure. This makes MG23 a novel and timely target for further in-depth study to unravel the molecular role played by its reported oligomerization and provide a better understanding of its potential druggability. Although our ability to study oligomerization of plasma membrane integral proteins has greatly advanced over the last few years, these approaches rely on extreme optical sectioning, which is not amenable to internal organelles such as the ER/SR. The aim of this PhD project is to develop and apply spatial-temporal molecular brightness approaches using confocal microscopy to address the problem of the oligomerization dynamics of MG23 in pathophysiological conditions. The use of fluorescence fluctuation spectroscopy offers promise to study homo-oligomerization in crowded systems, where the visualization of individual molecules is challenged either by protein density and/or by the local geometry of the sample. Using a combination of molecular biology, electrophysiology, heart failure models and advanced microscopy techniques, I will visualize MG23 functional oligomerization in cardiac cells, unravel MG23 oligomerization in heart failure conditions, and correlate MG23 oligomerization to cardiac cell pathology, specifically apoptosis and fibrosis. This project will develop a toolbox to study ER membrane protein oligomerization, with ramifications to the broader study of the unfolded protein response pathway in heart failure.

内质网（ER）稳态正在成为心力衰竭中越来越令人感兴趣的目标，因为错误折叠或未折叠蛋白质的积累很容易导致内质网应激，进而在心血管疾病的发展中发挥重要作用。内质网跨膜蛋白可以发挥影响细胞死亡或存活的重要信号作用。 Mitsugumin 23 (MG23) 是一种新发现的内质/肌浆网 (ER/SR) 跨膜蛋白，被认为本质上是寡聚体。我们认为 MG23 的动态寡聚行为控制其功能作用。我们的最新数据表明 MG23 在心脏纤维化、SR Ca2+ 渗漏和心力衰竭早期阶段中发挥作用。这使得 MG23 成为进一步深入研究的新颖而及时的靶标，以揭示其报道的寡聚化所发挥的分子作用，并更好地了解其潜在的成药性。尽管我们研究质膜整合蛋白寡聚化的能力在过去几年中已经大大提高，但这些方法依赖于极端的光学切片，这不适合 ER/SR 等内部细胞器。该博士项目的目的是开发和应用使用共焦显微镜的时空分子亮度方法来解决病理生理条件下 MG23 的寡聚动力学问题。荧光涨落光谱的使用为研究拥挤系统中的同源寡聚化提供了希望，其中单个分子的可视化受到蛋白质密度和/或样品的局部几何形状的挑战。结合分子生物学、电生理学、心力衰竭模型和先进的显微镜技术，我将可视化心肌细胞中的 MG23 功能性寡聚化，解开心力衰竭条件下的 MG23 寡聚化，并将 MG23 寡聚化与心肌细胞病理学（特别是细胞凋亡和纤维化）相关联。该项目将开发一个工具箱来研究内质网膜蛋白寡聚化，从而对心力衰竭中未折叠蛋白反应途径进行更广泛的研究。