Oxidative phosphorylation is the process by which mitochondria produce ATP (adenosine triphosphate) in the cellular respiratory chain, which is a key step in cellular energy metabolism. In this process, electrons are transferred from one carrier to another through a series of redox reactions and eventually to oxygen, forming water. The energy generated by this electron transport process is used to transfer the phosphate group to ADP (adenosine diphosphate) to produce ATP.
1. Detection method:
1.Spectrometry: Assess the efficiency of oxidative phosphorylation by measuring changes in the absorption or fluorescence spectrum of certain components in the mitochondrial respiratory chain.
2.Radiolabeling: Using radiolabeled phosphate, oxidative phosphorylation is assessed by measuring the extent to which it is embedded in ATP.
3.ATP yield assay: Direct measurement of the amount of ATP produced in cell or mitochondrial preparation. This can be achieved through various biochemical methods, such as the use of ATP-dependent enzymatic reactions.
4.Oxygen consumption rate (OCR) assay: Measure the oxygen consumption rate of cells using equipment such as the Seahorse XF analyzer to indirectly assess the efficiency of oxidative phosphorylation.
5.Fluorescent probes: Specific fluorescent probes are used to monitor changes in mitochondrial membrane potential, which are closely related to the efficiency of oxidative phosphorylation.
2. Application:
1.Bioenergetics research: Understand the fundamentals of cellular energy metabolism. 2.Disease mechanisms: to study alterations in oxidative phosphorylation in various diseases (e.g., neurodegenerative diseases, cancer). 3.Drug Screening & Toxicity Testing: Assesses the effect of drugs on mitochondrial function. 3. Precautions:
1.Sample Handling: Precise sample handling is required to maintain mitochondrial integrity. 2.Experimental conditions: Experimental conditions (e.g., temperature, oxygen concentration) have a significant impact on the results. 
Figure 1Phosphorylation quantitative proteomics analysis workflow.