Boosts cellular metabolism
Restores pH balance.
Repairs damaged cells
Produces more efficient protein receptors.
Provides proteins with crucial minerals
Mitochondria uptake and cellular respiration
Mitochondria are responsible for cellular respiration and energy production within a cell. Their growth and production are dependent on protein uptake. The proteins that interact with the mitochondria are typically produced via ribosome molecules within the cytosol. Interestingly, only proteins that are already translated or synthesized from RNA are permitted to enter the mitochondrial membrane and are not typically synthesized inside the mitochondria even though the mitochondria contain protein synthesizing compounds including DNA and RNA 1 .
The proteins, that pass through the mitochondrial membrane are composed of amino acids that are both hydrophobic (non-polar water repelling) and hydrophilic (dissolved in water due to charge interactions). After the protein is synthesized, it will try to fold and twist into the lowest energy state. Protein folding, results from the various hydrophobic amino acids within the chain being pushed away and compacted into the smallest geometric structure possible. This reduction of surface area will ensure that only very few amino acid molecules interact with the water surrounding water and any water present within the amino acids. The hydrophilic amino acids will try to maintain proximity to the water so that ion exchange can be readily undergone. Protein folding is stabilized by hydrogen bonding between the amino acids and a correctly folded protein is essential for it to function properly 2 .
There are protein chaperone molecules that prevent the protein from folding into its final 3D functional structure, the chaperones also guide the proteins close to the membrane where it can complete its ultimate destination as a folded 3D structure. Even though most folding is prevented outside of the membrane, there is a small section of protein that does fold and yields a charge gradient. The charge gradient is produced from a region of positive charges that will be oriented towards outer edges and the negative charges are tucked inside. This is commonly referred to an alpha helix 1 and is pictured below in figure 1. This helical section of protein is labeled the signal sequence and required for entry into the mitochondria.
The signal sequence interacts with a protein receptor complex that is located on the outer membrane of the mitochondria. The complex (TOM) forms a circular transfer channel that allows the protein and small ions to pass through the membrane. The electromagnetic forces from the helix, interact with the polar forces of the TOM complex and move the signal complex through the channel. A similar complex called the (TIM) complex, has an alpha helix tail connecting the inner and outer membranes. The tail can interact with the signal helix and through electron transfer, move the protein down the tail of the complex. A simple comparative analogy would be to consider a wire that must be snaked through a conduit. The wire is attached to the snake and moved gently through the conduit to eliminate snags. The conduit represents the transfer channel and the helix tail represents the snake. Both protein complexes permit membrane transfer, however the complexes typically work in conjunction 1 with one another. The TOM receptor brings the signaling helix into the intermembrane space, and as a result, the rest of the protein is in proximity to the TIM translocation tail and can interact with the tail. A cartoon image of the TIM and TOM complexes are pictured below in figure 2.
Figure 1. The red color represents positively charged amino acids. The yellow represents negatively charged amino acids that are tucked inside the helix.
Upon entry into the inner membrane or mitochondrial matrix space, the signal proteins undergo a terminal separation where the helical signal section is cleaved, and the remainder of the protein is filtered through to the inner membrane inside the matrix space. After the signal complex has been cleaved, the protein will fold into its 3D structure and is now functionally activated. A graphic representing the protein translocation process is pictured below in figure 3. It has been determined, that for the signaling helix, a specific amino acid sequence is not required, but rather only requires the formation of a polar gradient helix 1 . Due to this, a designed or engineered helical terminal sequence could be used to introduce any protein like molecules into the mitochondria.
Figure 2. The left side of the image illustrates a helical protein complex tail that is used to interact with the signal helix. The right side of the images represents a TOM complex that filters the signal protein through a translocation channel.
Figure 3. A graphic representing how proteins are permitted to enter in to the mitochondria matrix. A protein is recognized by receptors, that allow the protein to inter the matrix. After entry the protein undergoes protein folding and is biologically functionalized.
Transport between the mitochondria and the rest of the cell is dependent on energy production. Energy can be produced by hydrolysis, which is a process where molecules are broken down with water, and that break down releases and produces energy, often in the form of heat. One example of cellular energy is seen from adenine triphosphate molecule (ATP). Adenine is one of the four constituent nucleobases in DNA and RNA and is present in large quantities throughout an organism. The adenine is attached to a ribose sugar and then three phosphate groups are connected to the ribose. The triphosphate group is energetically unfavorable, due to the three negative charges 3 that are associated with the oxygen molecules. A graphic depicting ATP is pictured below in figure 4. The large phosphor molecules force the proton deficient oxygens close together and their negative charges are quick to repel. Thus, during hydrolysis, ATP will energetically remove the most accessible phosphate which is the gamma phosphate. The gamma phosphate typically has two charge locations that will quickly move toward a more positive charge and repel from the negative charge next to it. A hydroxyl group from a water molecule will replace the gamma phosphate. This replacement will result in a charge that is more balanced or neutralized and is termed adenine diphosphate (ADP). Energy will be produced as a result of this ion exchange and that same energy is stored by ADP until it is required for other reactions.
Figure 4. A representation of ATP illustrating negative oxygen charges. The gamma phosphate is the most accessible and has the largest negativity. When hydrolysis occurs, water replaces and gamma phosphate and may also quench the negative charge associate with the beta and alpha phosphate groups.
ATP is produced through cellular respiration. Cellular respiration occurs when oxygen breaks down glucose molecules and produces nicotinamide adenine dinucleotide (NAD) or flavin adenine dinucleotide (FAD) in a process called glycolysis. If there is not enough oxygen present, glycolysis become hindered and then, an energy shortage limits protein functionality. The dinucleotides produced are enzymes that gather up electrons and protons by acting as electron acceptors. They participate in a reduction reactions 9 as part of the glucose breakdown and become protonated in the form NADH or FADH2. The enzymes move toward the mitochondrial membrane and are stripped of the electrons they previously acquired. The stripping and translocation of protons is known as the electron transport chain. The inner membrane of a mitochondria is does not allow all molecule to penetrate through its channels, but it is very easy for electrons to traverse the membrane.
The embedded enzymes that are part of the electron transport chain, are known as shuttle enzymes 7 and allow protons to traverse the mitochondrial membrane. As an excess of protons begins to accumulate there is a need for regulation. The regulator is known as ATP synthase, and consumes the excess protons, it also produces a 3x amount of ATP as there were NADH molecules. It does this by using the protons and energetically forcing ADP from a diphosphate back to a triphosphate 3,4,6 . This process is pictured below in figure 5. Energy produced from ATP is required for protein complex binding inside the membrane or allows chaperones to interact with proteins moving towards the membrane. The energy is also used to initiate protein folding inside the membrane and break down glucose for cellular respiration. ATP is essential for cellular respiration, and without it there would not be enough energy for the cell to function properly.
Figure 5. A graphic depicting an electron transfer chain on the left side. A representation of ATP producing complex ATP synthase on the right. The protons enter in to FO domain generating a torque in the stalk where the F1 domain can convert ADP to ATP. This will ultimately produce the energy required to break down other glucose molecules and continue the cycle.
All these processes must function properly to regulate biological functions. If there is an excess of charge, electrolytes can be useful for balance and regulation. Electrolytes control how much water is present within the cellular environment. If the body is lacking in water, the cellular respiration will not have the protons needed to associate with NAD and then be consumed in ATP production. As we saw previously a reduction in ATP will result in considerably less energy for the whole organism. Also, many ionic elements are required and essential for proper protein function 9 inside the membrane. The protein binds to a separate cofactor enzyme and then may require Iron, Calcium, Magnesium, Potassium or other vital ions to assist in catalytic reactions. This is a regulatory feature, that requires a protein be enabled in order to accomplish it proper function, i.e. an on/off switch. Our product, Mito Restore, acts as a chelator and electrolyte. The chelation gathers free metal ions and collects them in one place or transports them near the mitochondria. Where they can be used for protein activation. It also binds other stronger metals that are not essential for protein interaction.
The main driver of the electrolyte solution is a small molecular weight carbon molecule that is easily broken down via glycolysis and hydrolysis. Mito restore is readily consumed and broken down into its constituent parts which ultimately resemble molecules like glucose, protein complexes, and to some extent, a helical amino acid chain. This means that Mito Restore has multifold benefits, for example, it acts a transport to supply the mitochondria with ions. These ions can easily traverse the membrane via the ion transport channels that allow protein entry and bind with the catalytic enzymes. It can also maintain and balance the pH level of the cellular environment. This balance of pH is crucial in the ATP production that produces the energy required for the cofactor enzymes to pair with mitochondrial proteins and help them to function properly. The pH balance is provided by the electrolyte solution in Mito Restore acting as either a proton donor or a proton receiver 10 . It can also act as a conductor of electricity, meaning that if there is a lack of electrons, the conductor can move electrons easily across the double bonded carbons present in the Mito Restore Solution. A conductive solution that allows electrons to move locations can increase the efficiency of the electron transport chain or restore balance to a transport chain that is malfunctioning.
After the Mito Restore participates in either glycolysis or hydrolysis and is broken down into its constituent molecules, they can access the internal membrane with the help of the membrane transport proteins mentioned above. The resemblance to glucose can allow a further breakdown that mimics the cellular respiration cycle of producing NAD from glucose. It also oxygenates the cellular environment. Thus, if used correctly Mito Restore can increase both cellular energy and metabolism by providing a bountiful supply of carbon and oxygen that is used and consumed during the metabolic process. As we saw previously, when a cell is deprived of oxygen the glycolysis process is hindered, and if there is insufficient supply of glucose for the oxygen to break down, then glycolysis and cellular respiration will not initiate.
1. Alberts B, Johnson A, Lewis J, et al. Molecular Biology of the Cell. 4th edition. New York: Garland Science; 2002. The Transport of Proteins into Mitochondria and Chloroplasts. Available from: https://www.ncbi.nlm.nih.gov/books/NBK26828/
2. Retrieved April 15, from https://www.ius.edu.ba/sites/default/files/u796/protein_folding_notes.pdf
3. ATP and reaction coupling. (n.d.). Retrieved from https://www.khanacademy.org/science/biology/energy-and-enzymes/atp-reactioncoupling/a/atp-and-reaction-coupling Bionomic Solutions
4. Berg JM, Tymoczko JL, Stryer L. Biochemistry. 5th edition. New York: W H Freeman; 2002. Section 18.5, Many Shuttles Allow Movement Across the Mitochondrial Membranes. Available from: https://www.ncbi.nlm.nih.gov/books/NBK22470/
5. Tortora, G. J., Funke, B. R., & Case, C. L. (2013). Microbiology: An Introduction. 13 edition 2014. San Francisco, CA: Benjamin-Cummings Publishing Company. Ch 27 Fluids, Electrolytes Acid base balance
6. Chemiosmosis and ATP Synthase. (2009). Retrieved from http://faculty.ccbcmd.edu/~gkaiser/biotutorials/energy/atpsynthase_il.html
7. Jonckheere, A. I., Smeitink, J. A., & Rodenburg, R. J. (2011). Mitochondrial ATP synthase: architecture, function and pathology. Journal of Inherited Metabolic Disease, 35(2), 211-225. doi:10.1007/s10545-011-9382-9
8. Rassow, J., & Pfanner, N. (2000). The Protein Import Machinery of the Mitochondrial Membranes. Traffic, 1(6), 457-464. doi:10.1034/j.1600-0854.2000.010603.x
9. Stein, L. R., & Imai, S. (2012). The dynamic regulation of NAD metabolism in mitochondria. Trends in Endocrinology & Metabolism, 23(9), 420-428. doi:10.1016/j.tem.2012.06.005