Roede James R, Go Young-Mi, Jones Dean P
Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado, Aurora, CO, USA.
Division of Pulmonary, Allergy and Critical Care Medicine, Department of Medicine, Emory University, Atlanta, GA, USA.
Methods Mol Biol. 2018;1782:197-227. doi: 10.1007/978-1-4939-7831-1_12.
Mitochondrial energy metabolism depends upon high-flux and low-flux electron transfer pathways. The former provide the energy to support chemiosmotic coupling for oxidative phosphorylation. The latter provide mechanisms for signaling and control of mitochondrial functions. Few practical methods are available to measure rates of individual mitochondrial electron transfer reactions; however, a number of approaches are available to measure steady-state redox potentials (E ) of donor/acceptor couples, and these can be used to gain insight into rate controlling reactions as well as mitochondrial bioenergetics. Redox changes within the respiratory electron transfer pathway are quantified by optical spectroscopy and measurement of changes in autofluorescence. Low-flux pathways involving thiol/disulfide redox couples are measured by redox Western blot and mass spectrometry-based redox proteomics. Together, the approaches provide the opportunity to develop integrated systems biology descriptions of mitochondrial redox signaling and control mechanisms.
线粒体能量代谢依赖于高通量和低通量电子传递途径。前者提供能量以支持氧化磷酸化的化学渗透偶联。后者提供线粒体功能的信号传导和控制机制。目前几乎没有实用的方法来测量单个线粒体电子传递反应的速率;然而,有多种方法可用于测量供体/受体对的稳态氧化还原电位(E),这些方法可用于深入了解速率控制反应以及线粒体生物能量学。呼吸电子传递途径中的氧化还原变化通过光谱学和自发荧光变化的测量来量化。涉及硫醇/二硫键氧化还原对的低通量途径通过氧化还原蛋白质免疫印迹和基于质谱的氧化还原蛋白质组学进行测量。这些方法共同为开发线粒体氧化还原信号传导和控制机制的综合系统生物学描述提供了机会。