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The mitochondrial respiratory chain on the inner mitochondrial membrane (IMM) is the main intracellular source of reactive oxygen species (ROS). Mitochondria are continuously exposed to ROS and accumulate oxidative damage faster than other parts of the cell. Many studies have linked ROS-induced mitochondrial dysfunction to necrotic and apoptotic cell death.
Fig. 1 Molecular structure of the two mito-targeted antioxidants and their situation into the inner mitochondrial membrane and electron transport chain. (Cantarero et al., 2020)
The most effective way to inhibit ROS-induced cytochrome C release and cell death is to prevent lipid peroxidation in the IMM.
However, there is no available antioxidant that specifically targets mitochondria, let alone IMM. in addition, most antioxidants have poor cellular permeability and require concentrations greater than 100 μm to prevent oxidative cell death.
As a leader in the field of cellular stress, Creative Bioarray can help our customers develop a range of peptide antioxidants that are taken up by mitochondria and concentrated in the IMM.
We can help our customers develop a new class of small cell permeable peptide antioxidants targeting mitochondria. The basic structural motif of these peptides consists of alternating aromatic residues and basic amino acids. These peptides contain a sequence of amino acids that are free to penetrate the cell despite carrying a net 3+ charge at physiological pH, and they are highly concentrated in the mitochondria.
We will customize the appropriate synthesis according to the customer's objectives.
The ability of targeted antioxidants to scavenge H2O2 in vitro is measured using luminol chemiluminescence assay. The antioxidant properties are further validated by inhibition of fatty acid peroxidation and low-density lipoprotein (LDL) oxidation.
To verify the uptake of targeted antioxidants by isolated mitochondria. Three cycles of freeze-thaw treatment were used to separate the inner and outer membranes to determine the localization of the peptide within the mitochondria.
Our evaluation of candidate peptide antioxidants includes, but is not limited to, cellular uptake and intracellular localization of peptide antioxidants, intracellular ROS and cell viability, intracellular mitochondrial potential, mitochondrial H2O2 production, mitochondrial oxygen consumption, mitochondrial membrane potential, mitochondrial swelling assay, mitochondrial cytochrome C release, animal model based studies.
Creative Bioarray is dedicated to providing high-quality products, comprehensive services, and tailored solutions to support and facilitate life sciences and pharmaceutical research and development. If you have any questions or needs, please contact us, and our customer service staff will help you the first time.