一个的分子工厂: 哺乳动物呼吸链的拆除和重建
Lukas Rimle1, Ben P Phillips2, Isabela M Codo Costa Barra2
1Department of Chemistry, Biochemistry and Pharmaceutical Sciences, University of Bern, Bern 3012, Switzerland.
概括
研究人员开发了一种可扩展的方法来净化和重组哺乳动物线粒体呼吸链复合体I,III,IV和ATP合成酶 (复合体V) 进入脂质体. 这种系统成功地证明了氧化酸化,并揭示了心血管脂蛋白.
科学领域:
- 生物化学 生物化学
- 细胞生物学 细胞生物学
- 结构生物学 结构生物学
背景情况:
- 哺乳动物线粒体呼吸复合体I-V对于ATP生产至关重要.
- 单个复合物的体外研究是常见的,但在脂质体中多种哺乳动物复合物的共同重建是不太了解的.
- 了解功能组合是描述生物能源过程的关键.
研究的目的:
- 开发一种通用和可扩展的策略,用于净化和重组哺乳动物线粒体呼吸复合体I,III和ATP合成酶到脂质体中.
- 为了证明整个哺乳动物呼吸链 (复合体I,III,IV和V) 在脂质体中的功能共同复合.
- 为了研究心脏脂蛋白在激活哺乳动物呼吸链中的作用.
主要方法:
- 从丰富的线粒体中净化哺乳动物呼吸复合体I,III和ATP合成酶.
- 净化复合物的重新组装成脂质体.
- 在现场监测ATP合成速率和抑制研究.
- 电子冷显微镜 (cryo-EM) 用于结构特征.
主要成果:
- 纯化了酶活性的哺乳动物呼吸复合体I,III和ATP合成酶.
- 在脂质体中重建了一个功能性的哺乳动物呼吸链 (I,III,IV和V复合体),证明了NADH驱动的ATP合成.
- 卡迪奥利平被确定为激活哺乳动物呼吸链的重要组成部分.
- 化电磁场提供了阿波状态复合物III的高分辨率结构和复合物V的中分辨率结构.
结论:
- 建立了一种可扩展的方法,用于将哺乳动物呼吸系统复合体共同重构成功能性蛋白质体.
- 重建后的系统可以研究氧化酸化和脂质的作用,如心脏脂蛋白.
- 这项工作为未来的线粒体生物能学的表征和调制提供了基础.
相关概念视频
01:41The Supercomplexes in the Crista Membrane
2.5K
The mitochondrial cristae membrane is the primary site for the oxidative phosphorylation (OXPHOS) process of energy conversion mediated through respiratory complexes I to V. These complexes have been widely studied for decades, and it has been proven that they form supramolecular structures called respiratory supercomplexes (SC). These higher-order complexes may be crucial in maintaining the biochemical structure and improving the physiological activity of the individual complexes while...
2.5K
01:28Electron Transport Chains
96.7K
The final stage of cellular respiration is oxidative phosphorylation that consists of two steps: the electron transport chain and chemiosmosis. The electron transport chain is a set of proteins found in the inner mitochondrial membrane in eukaryotic cells. Its primary function is to establish a proton gradient that can be used during chemiosmosis to produce ATP and generate electron carriers, such as NAD+ and FAD, that are used in glycolysis and the citric acid cycle.
The ETC is comprised of...
The ETC is comprised of...
96.7K
01:30The Electron Transport Chain
15.9K
The electron transport chain or oxidative phosphorylation is an exothermic process in which free energy released during electron transfer reactions is coupled to ATP synthesis. This process is a significant source of energy in aerobic cells, and therefore inhibitors of the electron transport chain can be detrimental to the cell's metabolic processes.
Inhibitors of the electron transport chain
Rotenone, a widely used pesticide, prevents electron transfer from Fe-S cluster to ubiquinone or Q...
Inhibitors of the electron transport chain
Rotenone, a widely used pesticide, prevents electron transfer from Fe-S cluster to ubiquinone or Q...
15.9K
01:46Electron Transport Chain: Complex I and II
9.9K
The mitochondrial electron transport chain (ETC) is the main energy generation system in the eukaryotic cells. However, mitochondria also produce cytotoxic reactive oxygen species (ROS) due to the large electron flow during oxidative phosphorylation. While Complex I is one of the primary sources of superoxide radicals, ROS production by Complex II is uncommon and may only be observed in cancer cells with mutated complexes.
ROS generation is regulated and maintained at moderate levels necessary...
ROS generation is regulated and maintained at moderate levels necessary...
9.9K
01:43Electron Transport Chain: Complex III and IV
6.8K
During the electron transport chain, electrons from NADH and FADH2 are first transferred to complexes I and II, respectively. These two complexes then transfer the electrons to ubiquinol, which carries them further to complex III. Complex III passes the electrons across the intermembrane space to Cyt c, which carries them further to complex IV. Complex IV donates electrons to oxygen and reduces it to water. As electrons pass through complexes I, III, and IV, the energy released aids the pumping...
6.8K
01:22Introduction to Cellular Respiration
172.2K
Organisms harvest energy from food, but this energy cannot be directly used by cells. Cells convert the energy stored in nutrients into a more usable form: adenosine triphosphate (ATP).
ATP stores energy in chemical bonds that can be quickly released when needed. Cells produce energy in the form of ATP through the process of cellular respiration. Although much of the energy from cellular respiration is released as heat, some of it is used to make ATP.
During cellular respiration, several...
ATP stores energy in chemical bonds that can be quickly released when needed. Cells produce energy in the form of ATP through the process of cellular respiration. Although much of the energy from cellular respiration is released as heat, some of it is used to make ATP.
During cellular respiration, several...
172.2K