Dissecting the Dynamic Landscape of TCA-Cycle Enzyme Interactions in Plant Mitochondria

三羧酸循环是细胞代谢的重要代谢中枢，对线粒体ATP的产生至关重要。在植物中，它受到白天光合作用活动和夜间不光合作用引起的功能振荡的影响。目前还不完全清楚这种功能性振荡是如何调节的。本项目的目的是阐明三羧酸循环中酶的相互作用变化是否对三羧酸循环具有调节作用。为此，利用凝胶电泳和质谱分析方法分析了三羧酸循环蛋白的相互作用，从而可以深入了解这些酶的丰度和组成。在第一步，两种已经存在的蛋白质分析方法：蛋白质相互作用将被结合起来。这不仅将减少过程中蛋白质组装的人工降解，而且还提供结构见解。第二步，定量研究白天和夜间三羧酸循环酶之间蛋白质相互作用的差异。该项目的第三部分旨在确定导致蛋白质-蛋白质相互作用变化的机制。

The tricarboxylic acid cycle is an important metabolic hub of cellular metabolism and essential for mitochondrial ATP production. In plants, it is subject to a functional oscillation caused by photosynthetic activity during the day and its absence during the night. It is currently not fully understood how this functional oscillation is regulated. The aim of this project is to clarify whether changing interactions of enzymes in the tricarboxylic acid cycle have a regulatory influence on the tricarboxylic acid cycle. For this purpose, the interactions of tricarboxylic acid cycle proteins are analyzed using gel electrophoresis and mass spectrometry based methods, which enables insight into abundance and composition of assemblies of these enzymes. In a first step, two already existing approaches for the analysis of protein:protein interactions will be combined. This will not only reduce artificial degradation of protein assemblies during the procedure, but also provide structural insights. In a second step, the differences between the protein interactions of tricarboxylic acid cycle enzymes during the day and during the night are investigated quantitatively. The third part of the project then aims at identifying the mechanisms that cause the changes in the protein-protein interactions.