Protein kinase D-mediated balance between lipid homeostasis and secretory activity in mammalian cells - A systemic appoach

蛋白激酶 D 介导的哺乳动物细胞脂质稳态和分泌活性之间的平衡 - 系统方法

• 批准号: 191645520
• 负责人: Privatdozentin Dr. Angelika Hausser
• 金额: --
• 依托单位: Institut für Zellbiologie und Immunologie (IZI)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/191645520?language=en
• 关键词: Protein; kinase; mediated; balance; between

Lipid metabolism, sorting and transport between different organelles are tightly regulated in mammalian cells, since lipid homeostasis is critical for maintaining the integrities of cellular membranes. At the Golgi complex the lipid composition determines the formation of transport vesicles, which are required to shuffle secretory proteins from the trans-Golgi network (TGN) to the cell membrane. Protein kinase D (PKD) has been shown to play an important role in this regulation: Active PKD is directly involved in the fission of transport vesicles at the TGN. Furthermore, it has a dual function for the ceramide transport from the endoplasmic reticulum (ER) to the TGN by regulating the activity of the ceramide transfer protein (CERT) in two opposite ways. PKD and CERT mutually regulate each other in a complex structure of interrelated feedback circuits, making this system an attractive target for a holistic modeling approach. Moreover, recent advances in experimental techniques have led to new insights about molecular interactions at the TGN, and regulation of secretion has not yet been taken into account in theoretical secretion models. The goal of this project is to increase our understanding about the role of PKD in the balance between lipid homeostasis and secretion. This will be achieved by a data- and knowledge-driven modeling approach based on chemical reaction kinetics, which will be accompanied by perturbation experiments that are used for model identification. We will particularly focus on statistical methods for parameter estimation and model selection, identifiability analysis, and systems-theoretic methods to investigate coupled feedback. The results of our project will be highly relevant for biotechnological applications: We may reveal targets for optimization of mammalian host cell lines used for therapeutic protein production, where regulation processes at the TGN have recently been identified as a bottleneck in the secretory pathways.

在哺乳动物细胞中，脂质代谢，不同细胞器之间的分类和转运受到严格调节，因为脂质稳态对于维持细胞膜的完整性至关重要。在高尔基体络合物上，脂质组成决定了传输囊泡的形成，这是将分泌蛋白从反式高尔基网络（TGN）调整为细胞膜所必需的。蛋白激酶D（PKD）已被证明在此调节中起着重要作用：活性PKD直接参与TGN的转运囊泡的裂变。此外，它通过以两种相反的方式调节神经酰胺转移蛋白（CERT）的活性，具有从内质网（ER）到TGN的双重功能。 PKD和CERT在相互关联的反馈电路的复杂结构中相互调节，使该系统成为整体建模方法的有吸引力的目标。此外，实验技术的最新进展导致了有关TGN分子相互作用的新见解，并且在理论分泌模型中尚未考虑分泌的调节。该项目的目的是提高我们对PKD在脂质稳态和分泌之间平衡中的作用的理解。这将通过基于化学反应动力学的数据和知识驱动的建模方法来实现，该方法将伴随着用于模型识别的扰动实验。我们将特别关注参数估计和模型选择，可识别性分析以及系统理论方法的统计方法，以研究耦合反馈。我们项目的结果将与生物技术应用高度相关：我们可能会揭示用于优化用于治疗蛋白质生产的哺乳动物宿主细胞系的靶标，在这些哺乳动物的宿主细胞系中，最近已将TGN的调节过程确定为分泌途径中的瓶颈。