The assessment of individual enzymes involved in mitochondrial oxidative phosphorylation (OXPHOS) is an essential component of manifestation characterizing mitochondrial disease and can offer crucial insights into the functionality of mitochondrial OXPHOS across diverse cell types and tissues. Protheragen is a leading provider of specialized services for mitochondrial analysis. Leveraging multiple biochemical techniques, our team of experts efficiently and accurately conducts the determination and analysis of mitochondrial respiratory chain complexes as well as ATP enzymes.
Fig.1 Possible sites of ROS production (red arrows) in respiratory chain. (Mazat, J. P., et al., 2020)
Our Services
The services provided by Protheragen are tailored and comprehensive measurements of mitochondrial respiratory chain enzyme activity. Our service is built upon validated and dependable protocols for isolating mitochondria from diverse samples, as well as spectrophotometric determination of mitochondrial respiratory chain complexes I-V in addition to citrate synthase (CS).
Measurement of Complex I (NADH: Ubiquinone Oxidoreductase) Activity
Complex I serves as the initial site of the mitochondrial respiratory chain, facilitating the oxidation of NADH produced by the Krebs cycle in the mitochondrial matrix. It transfers two electrons to CoQ, an electron carrier, through flavin mononucleotide (FMN) and a series of iron-sulfur (Fe-S) clusters.
Measurement of Complex II (Succinate: Coenzyme Q Oxidoreductase) Activity
Complex II plays a role in converting succinate to fumarate during the sixth step of the TCA cycle within the mitochondrial matrix. This enzyme combines oxidation with CoQ's reduction, serving as an electron carrier.
Measurement of Complex III (Ubiquinol: Cytochrome c Oxidoreductase) Activity
Complex III, also known as cytochrome bc1 complex, facilitates electron transfer from ubiquinol to cytochrome c -the second mobile electron carrier in the mitochondrial respiratory chain.
Measurement of Complex IV (Cytochrome c Oxidase) Activity
Cytochrome c oxidase acts as a terminal enzyme within the electron transport chain (ETC). It catalyzes both oxidation reactions involving cytochrome c and reduction reactions converting molecular oxygen (O2) into water (H2O).
Measurement of Complex V (ATP Synthase) Activity
Complex V functions as an F-type ATP synthase. It operates as a rotary enzyme coupling proton translocation driven by proton motive force generated at complexes I, III, and IV with ATP synthesis.
Our Advantages
Protheragen is a leading provider of custom services in the field of Mitochondrial Respiratory Chain Enzyme Analysis, dedicated to delivering innovative solutions that drive your biological research forward. If you have any questions or require general information, please do not hesitate to contact us. We are always happy to assist you.
Reference
- Mazat, J. P.; et al. (2020). Modelling mitochondrial ROS production by the respiratory chain. Cellular and molecular life sciences: CMLS, 77(3), 455–465.
