Mitochondria, a crucial organelle involved in the generation and regulation of cellular bioenergetics, play a pivotal role in supplying energy in the form of ATP through the oxidative phosphorylation system (OXPHOS). Consequently, it is not surprising that mitochondrial dysfunction has been implicated in various pathophysiological processes and mitochondrial diseases. Protheragen takes pride in being a leading service provider offering comprehensive mitochondrial oxidative phosphorylation analysis worldwide.
Overview of the Mitochondrial Oxidative Phosphorylation System
The oxidative phosphorylation (OXPHOS) system comprises five protein complexes and two electron carriers that are embedded in the inner mitochondrial membrane. High-energy phosphate production is achieved by coupling electron transfer to proton translocation across the inner membrane, resulting in the establishment of an electrochemical gradient. This gradient generates a motive force that drives ATP synthesis through the fifth complex, ATP synthase (complex V [CoV]). During respiration, electrons originating from NADH and succinate, products of the citric acid cycle, are sequentially transferred through complexes I (CoI) and II (CoII), respectively, to ubiquinone. Subsequently, they pass through complex III (CoIII) and cytochrome c before finally reaching complex IV (CoIV).
Fig.1 Mitochondrial Oxidative Phosphorylation. (Purandare, N., et al., 2023)
Our Services
The analysis of oxidative phosphorylation and the function of the mitochondrial electron transfer system (ETS) can provide comprehensive information on metabolite uptake, metabolization, enzyme activities in the mitochondrial matrix, and the coupling of electron transfer with ADP phosphorylation to generate ATP. This reflects the mitochondria's ability to meet cellular energy demands. With various biochemical methods and advanced instruments, Protheragen efficiently and accurately detects changes in oxidative phosphorylation enzyme activity and oxygen consumption associated with mitochondrial diseases. Our platform specializes in detecting oxidative phosphorylation in mitochondrial disease.
Oxidative Phosphorylation Enzyme Activity Measurement Service
Our platform utilizes biochemical methods to efficiently and accurately detect alterations in the activity of enzymes related to oxidative phosphorylation. Currently, we are capable of detecting the following substances.
- Na+K+ ATP Enzyme activity
- Ca2+Mg2+ ATP Enzyme activity
- Activity of Complex I (NADH: ubiquinone oxidoreductase)
- Activity of Complex II (Succinate: coenzyme Q oxidoreductase)
- Activity of Complex III (Ubiquinol: cytochrome c oxidoreductase)
- Activity of Complex IV (Cytochrome c oxidase)
- Activity of Complex V (ATP synthase)
- Measurement of Oxygen Consumption Rate (OCR)
As a pivotal indicator for mitochondrial function, OCR is employed to investigate the functionality of mitochondrial oxidative phosphorylation. The assay typically measures basal respiration under normal conditions and subsequently introduces oligomycin to inhibit ATP synthase, resulting in a significant decrease in OCR. This leaves only the oxygen consumption rate attributable to proton leak. The remaining portion represents the oxygen consumption rate associated with oxidative phosphorylation (ATP production).
- Assay for ATP Content
The platform we utilize employs a robust biochemical assay to accurately quantify ATP levels, thereby elucidating alterations in cellular energetics.
Our Advantages
The team at Protheragen is dedicated to delivering professional mitochondrial oxidative phosphorylation analysis for mitochondrial diseases. If you are interested in availing our services, please feel free to contact us.
Reference
- Purandare, N.; et al. (2023). Mitochondrial Oxidative Phosphorylation in Viral Infections. Viruses, 15(12), 2380.
