SUPEROXIDE DISMUTASES (SOD) AND NUTRITIONAL MODULATION AGAINST OXYGEN FREE RADICALS - Pt. 2

The hydrogen peroxide produced by the action of superoxide dismutses and other oxidative processes can be detoxified by two different systems.  The enzyme catalase converts hydrogen peroxide to water.

             2H₂O₂  --> 2H₂O  +  O₂           (2)

This reduction of hydrogen peroxide by catalase occurs without the intermediate formation of the hydroxyl radical.  Catalase is a homoprotein that is found almost exclusively within peroxisomes.  Similar to the inhibition of superoxide dismutase, inhibition of catalase with aminotriazole potentiates oxidative stress cell injury.

The enzyme glutathione peroxidase also catalyzes the reduction of hydrogen peroxide to water.  Reduced glutathione is used as the source of reducing equivalents necessary to drive this reaction.  In this process, two molecules of glutathione are oxidized to yield one molecule of glutathione disulfide (GSSG).  Glutathione disulfide is efficiently reduced to glutathione by glutathione reductase.  NADPH serves as the source of the reducing equivalents.  The glutathione peroxidase-reductase system seems to be the first line of defense against hydrogen peroxide, and catalyse is a secondary system.  Catalase is also a specific defense against hydrogen peroxide formed within the peroxisomes.

Glutathione peroxidase is found in the cytosol of most cells as well as within the mitochondria.  Glutathione peroxidase also shows considerable activity toward organic hydroperoxides, converting them to their corresponding alcohols.  By contrast, catalase will only catabolize hydrogen peroxide.  The cytosolic and mitochondrial forms of glutathione peroxidase are dependent upon the metal selenium for activity.  It is not surprising that selenium deficiency has been demonstrated to exacerbate oxidant-stress injury ²³ and when selenium and vitamin E are depleted in a tumor cell the cell is carcinogenic.²⁴  It is very important to note that SODs within the cell may be found to be interdependent upon the fat soluble vitamin such as vitamin E vs. the water soluble vitamin C as found on the outer cell membrane.  Their common link being the phospholipidation of the cell, especially the skin cells where compounds may be absorbed within 25 seconds into the circulatory system vs. 10 seconds for dimethylsulfoxide (DMSO) and olive oil.²⁵

The importance of glutathione reductase is emphasized by the observation that inhibition of this enzyme also potentiates the cytotoxicity of oxidative stress.  With the inhibition of glutathione reductse, oxidized glutathione is not converted to reduced glutathione, a result that limits the effectiveness of the peroxidase by limiting the supply of glutathione.  The chemotherapeutic alkylating agent, 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) inhibits glutathione reductase without effect on glutathione peroxidase and catalase.  This compound sensitizes a number of cell types to the toxicity of hydrogen peroxide and other organic hydroperoxides.  It should also be noted that, during severe oxidative stress, NADPH levels are markedly depleted, and effect related to the consumption of NADPH by glutathione reductase.  Such a depletion may exert a substrate \-level inhibition of the reductase.  Thus, the NADPH depletion that results from oxidative stress may actually sensitize the cell to the oxidative stress.  Superoxide dimutase and catalase provide antioxidant protection by inhibiting the formation of the hydroxyl radical as found in free oxygen radicals.

The Future of Superoxid Dismutases and Nutritional Modulation

NASA principal investigator, Dr. Gloria Borgstahl, formerly of the University of Toledo, and now of the University of Nebaraska, has successfully crystallized E. coli manganese superoxide dismutase (MnSOD), antioxidant enzymes that are homologous to those found in cellular mitochondria, on the International Space Station during the period of December 2001 to April 2002.  Several of the MnSOD crystals grown on ISS were 80 times greater in crystal volume than earth-grown crystals.  Di spots to 1.26 resolutions were observed  providing significantly improved data obtained from crystals grown in earth laboratories.  An exciting result was that the MnSOD crystals grown on ISS were suitable for neutron studies and time-Laue studies-methods that require large, perfect crystals. With the neutron examples the researchers hope to be able to obtain the, never-before-seen, three-dimensional structure of the hydrogen’s on each amino acid of the protein and thereby be able to answer the unsolved questions concerning the source of these hydrogens in this reaction mechanism.  With the time-resolved Laue experiments, the team will be able to generate the superoxide substrate within the crystals with a laser pulse and thus film the “movie” of the enzyme converting it to the products peroxide and water.  The  enzyme MnSODs in the body is important, and in-depth study of their structure is not to the ability to understand their true function, but these experiments may lead to new therapeutics for the treatment of various degenerative diseases.¹⁸

Superoxide dismutases (SODs) are antioxidant metalloenzymes catalyzing the redox disproportion in the (dismutation) of superoxide radical, O₂*- as previously discussed in Equation (1).

It is generally accepted that in all SODs the metal ion (M) catalyzes dismutation of the superoxide radical through a cyclic oxidation reduction mechanism:

           M³⁺  +  O₂^*-  -->  M²⁺  +  O₂       (1.1)

          M²⁺  +  O₂^*-  +  2H⁺  --->  M³⁺  +  H₂O₂    (1.2)

The four classes of SODs are known, distinguished by the metal prosthetic groups:  Cu/Zn, FE, Mn and Ni.   Fe^- and Mn^- SODs constitute a structural family.^26,27  Fe- and Mn- SODs are unequally distributed throughout the kingdoms of living organisms and are located in different cellular compartments.^28,29,30,31  In particular, Fe-SOD is found in obligate anaerobes and aerobic diazotrophs (exclusively), facultative aerobes (exclusively or together with Mn-SOD).  In the cytosol of cyanobacteria, in the chloroplast stroma of higher plants, in the protozoa, kelp, Yamatoshinjo and Oaky Smoky (Cu/Zn-SOD, Fe-SOD, Mn-SOD)^{TM 32,33,34}  Fe-SOD and Mn-SOD form some organisms (e.g. Escherichia coli ) exhibit almost absolute metal specificity ,³⁵ while other enzymes, such as “cambialistic” SOD form Propionibacterium shermanii, are captive with either metal.³⁶  Fe- and Mn-SODs occur as homodimers or homotetramiers. 

The 3-D structures of several Fe-SODs have been determined. ^{36,37,38,39,40}  The monomers fold into two domains.  The N-terminal domain consists of two long antiparallel helices.  The C-terminal domain contains a central beta-sheet formed by three antiparallel beta strands with 4-6 surrounding helices.  The iron atom is lignaded by two residues from each of the N-terminal helices and two residues from the loops in the C-terminal domain.

The active site iron is pentacoordinate, with the metal lignands (N ^{x epsilon}) of three conserved His residues, O ^{x delta} of the conserved Asp residue of a water molecule) arranged in distorted trigonal bipyramidal geometry, which opposite of the tetrahydronal molecule of water.  The first His residue and a solvent molecule fill the two axial positions.  In the azide-Fe^III –SOD complex, the iron becomes hexacoodinate with distorted octahedral geometry (similar to the fat soluble Vitamin E), with azide coordinated trans to ASP ligand.⁴¹ Table 1 lists the mononuclear iron environment residues in known 3D structures with their PDB Code and Reference.

Superoxide dismutases (SODs) have been found in various nutritional sources such as Jerusalem artichoke powder and Bifidobacterium. ^42,43 Juice Plus + Vineyard Blend, foods grown in red clay soils, Yamatoshinjo and hemp oil. ^{44, 34,45}  The use of these nutritional modulators have shown significant risk to oxidative stress via the superoxide radical as they have maintained cellular integrity and reduced cellular sensitization as individual whole foods with an individual’s meals.

Summary

Cells are continuously exposed to a variety of oxidative process which could potentially lead to cellular injury or death.  As a result, aerobic organisms possess effective defense mechanisms against oxidative stress as associated with the superoxide radical and improper nutritional modulation that would reduce superoxide dismutases (SODs) in the cell and thereby circumvent toxic cell injury.  These protective mechanisms fall into two broad categories:  (1) those which prevent the initiation of lipid peroxidation and (2) those which prevent its propagation.

Superoxide dismutase and catalase provide antioxidant protection by inhibiting the formation of the hydroxyl radical.  Chelation of the ferric iron necessary for the formation of the hydroxyl radical and direct removal of this species by radical scavengers (e.g., mannital) are also protective.  The generation of species capable of initiating peroxidation may exceed the capacity of effectively remove them.  Thus, there are other defenses to prevent the uncontrolled propagation of lipid peroxidation and oxidative stress as associated with free oxygen radicals/hydroxyl radicals.  These include water-soluble antioxidants, such as ascorbic acid (vitamin C) and reduced glutathione, and fat-soluble antioxidants, most notably alpha-tocopherol (vitamin E).  It appears that vitamins E and C and glutathione combine in some as yet poorly defined cycle to donate hydrogen atoms to lipid and/or peroxy radicals, thereby preventing further propagation of the lipid peroxidation, which was initiated by free oxygen radicals. 

The metabolism of toxic substances as associated with hazardous materials by mixed function oxidation or other mechanisms leads to irreversible cell injury thorough mechanisms that have been related either to the covalent binding of reactive metabolites, to changes in protein thiols, to alterations in intracellular calcium homeostasis, or to the formation of partially reduced oxygen species.  The relative roles that each of these mechanisms plays in any particular example of toxic cell injury remains controversial and a subject of continuing investigation through the advancement of new analytical technologies/equipment.  In the case of the various forms of superoxide dismutases, “quickened” advances in science, chemistry and wave genetics ⁴⁶ will show that these metalloenzymes will play a major character role in the explanation of “creation” through the variable of true far infrared sources, superluminal radiance, the effects of Cooper pairs upon Eccelerated^TM intelligent water and the role of the white worm hole found in black holes as recently found in the DNA molecule.  These future researchers will be the Indigo and Crystal children – our grand children with their little pet black rabbit with pink ears and a white spot  named Ben as it hops down a white worm hole in the quantum biophysics of the human cell.  Time and Eternity will show all of us the wonders that will amaze the original Creator of us all through the marvels of science, spirituality, art and medicine.

References

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  Volume 1 © 2004

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    © 2005

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   Orem, Utah. © 2004

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⁴⁵   Staninger, Hildegarde.  The Earthworm:  A Microcosmos of “No Disease” as Compared to the Human Being, A Macrocosmos of Chaos and the Disease, Cancer.  A Comparative Research Thesis.  Capital University of Integrative Medicine.  Washington, D.C., Nov. 4, 2001

⁴⁶   Staninger, Hildegarde.  Acoustical Wave Genetics.  Page One Science, Inc. Alexandria, VA

     © 2005.