Assessment of oxidant stress in vitro and in vivo

Details Assessment of oxidant stress in vitro and in vivo Assessment of oxidant stress in vitro and in vivo

2001-07-26

2004-11-02

Carlson, Karen Cochrane (Department: 1653)

Drug, bio-affecting and body treating compositions

Designated organic active ingredient containing

Peptide containing doai

Reexamination Certificate

active

06812212

ABSTRACT:

BACKGROUND OF INVENTION
1. Field of the Invention
The present invention generally relates to methods of assessing and preventing oxidant stress in vitro and in vivo. More specifically, the present invention relates to methods of measuring nitration and polymerization of proteins which can be used for assessing oxidant stress and methods of preventing nitration and/or polymerization of protein due to oxidant stress.
2. Description of Related Art
It is generally known in the art that superoxide is produced under oxidant stress. This reactive molecule directly or indirectly reacts with macromolecules such as DNAs and proteins and depletes reducing agents such as glutathione in the cells.
Prostaglandin H
2
synthases, which are membrane-bound enzymes, catalyze the committed step in the biosynthesis of the prostaglandins and thromboxanes. Prostaglandin H
2
synthases have cyclooxygenase and peroxidase activities (1). Prostaglandin H
2
synthases add molecular oxygen to arachidonic acid to form prostaglandin G
2
. Prostaglandin G
2
is then rapidly converted to prostaglandin H
2
by reduction of the peroxide to a hydroxyl group. Prostaglandin H
2
is enzymatically or non-enzymatically converted to prostaglandin F
2
&agr;, prostaglandin E
2
, prostaglandin D
2
, prostaglandin I
2
(prostacyclin) or thromboxane A
2
(2-4). In addition to constitutively expressed prostaglandin H
2
synthase form 1, a second inducible form of prostaglandin H
2
synthase, prostaglandin H
2
synthase form 2, was discovered in chicken fibroblasts (5) and murine 3T3 cells (6). Prostaglandin H
2
synthase form 2 is similar to prostaglandin H
2
synthase form 1 with regard to molecular size, subunit composition, and general reaction mechanism. The two prostaglandin H
2
synthase isoforms have 60% identity in their primary sequences (7).
Prostaglandin biosynthesis is the target for non-steroidal anti-inflammatory drugs (NSAIDs). Prostaglandin H
2
synthase is the primary target of aspirin, indomethacin, ibuprofen, and other NSAIDs (5,8). Increased expression of both inducible nitric oxide (NO) synthase and prostaglandin H
2
synthase form 2 has been reported in intestinal inflammation (9-11).
Cytochrome c induces apoptosis by translocation from the mitochondrial membrane to the cytoplasm followed by binding to the apoptotic protease activating factor-1 (Apaf-1) which activates caspases (12). Nitration and/or polymerization of cytochrome c in mitochondria may compromise its translocation and/or its caspase activation function.
Nitration of several amino acid residues of proteins is a result of the reaction of NO with superoxide which is produced under oxidant stress to form peroxynitrite, a potent toxic oxidant. Recently, it was reported that nitrotyrosine was also formed via a tyrosyl radical produced during catalysis of prostaglandin H
2
synthase form 2 (13). Nitration of caspase-3 (14), ribonucleotide reductase (15), and cytochrome P450 2B1 (16) resulted in loss of catalytic activity. Interestingly, nitration of cysteine residue of ovine prostaglandin H
2
synthase form 1 enhanced catalytic activity by alteration of secondary structure of the enzyme (17).
While it is well known in the art that oxidant stress can lead to further problems within the body which, as set forth above, there are no methods or products which limit or eliminate oxidant stress. These problems can include cancer and other ailments. Accordingly, it is desirable to develop new methods and markers for determining oxidant stress.
SUMMARY OF THE INVENTION
According to the present invention, there is provided a method of assessing oxidant stress by measuring polymerization of proteins. Also provided is a marker for oxidant stress which includes a polymerized protein. A kit for use in assessing oxidant stress, the kit including an assay for detecting polymerized proteins is also provided. A method of lowering oxidant stress by administering to a patient an effective amount of at least one reducing agent is also provided. A pharmaceutical composition for lowering oxidant stress, the pharmaceutical having an effective amount of reducing agent and a pharmaceutically acceptable carrier is also provided.


REFERENCES:
patent: 4029768 (1977-06-01), Penn
patent: 6133039 (2000-10-01), Heinecke
Czech et al. J Invest Dermatol. Jan. 1997: 108-12.*
Gadelha et al. Arch Biochem Biophys. Sep. 1997 15:345(2):243-50.*
Brown GC, Borutaite V. Nitric oxide, cytochrome c and mitochondria. Biochem Soc Symp. 1999;66:17-25. (Abstact).*
Cassina AM, Hodara R, Souza JM, Thomson L, Castro L, Ischiropoulos H, Freeman BA, Radi R. Cytochrome c nitration by peroxynitrite. J Biol Chem. Jul. 14, 2000;275(28):21409-15.*
Ahern H. The Scientist, 1995, vol. 9, No.: 15, p. 20.*
Smith, W.L. Marnett, L.J. Biochim. Biophys. Acta, 1083: 1-17 (1991).
Baker, R.R. Clin. Biochem., 23: 455-458 (1990).
Smith, W.L. Biochem. J., 259: 315-324 (1989).
Lote, C.J. Haylor, J. Prostags. Leukos. Essen. Fatty Acids, 36: 203-217 (1989).
Xei, W., Robertson, D.L. and Simmons, D.L. Drug. Develop. Res. 25: 249-265 (1992).
Kujubu, D., Fletcher, B., Varnum, B., Lim, R., & Hershman, H.R. J. Biol. Chem. 266:12866-12872 (1991).
Smith, W. L. and DeWitt, D. L. Advances in Immunol. 62: 167-215 (1996).
White, S. Prostagland. Leuk. Essent. Fatty, 43: 209-222 (1991).
Miller, M. J. S., Thompson, J. H., Zhang, X-J., Sadowskakrowicka, H., Kakkis, J. L., Munshi, U. K., Sandoval, M., Rossi, J. L., Elobychilress, S., Beckman, J. S., Ye, Y. Z., Rodi, C. P., Manning, P. T., Currie, M. G., and Clark, D. A. (1995) Gastroenterology 109, 1475-1483.
Reuter, B. K., Asfaha, S., Buret, A., Sharkey, K. A., and Wallace, J. L. (1996) J. Clin. Invest., 98, 2076-2085.
Singer, I. I., Kawka, D. W., Scott, S., Weinder, J. R., Mumford, R. A., Riehl, T. E., and Stenson, W. F. (1996) Gastroenterology 111, 871-885.
Green, D. R. and Reed, J. C. (1998) Science 281, 1309-1312 and references therein.
Gunther, M. R., Hsi, L. C., Curtis, J. F., Gierse, J. K., Marnett, L. J., Eiling, T. E., and Mason, R. P. (1997) J. Biol. Chem. 272, 17086-17090.
Rossig, L., Fichtlscherer, B., Breitschopf, K., Haendeler, J., Zeiher, A. M., Mulsch, A., and Dimmeler, S. (1999) J. Biol. Chem. 274,6828-6826.
Lepoivre, M., Fieschi, F., Coves, J., Thelander, L., Fontecave, M. (1991) Biochem. Biophys. Res. Commun. 179, 442-448.
Roberts, E. S., Lin, H-L., Crowley, J. R., Vuletich, J. L., Osawa, Y., and Hollenberg, P. F. (1998) Chem. Res. Toxicol. 11, 1067-1074.
Hajjar, D. P., Lander, H. M., Pearce, S. F. A., Upmacis, R. K., and Pomerantz, K. B. (1995) J. Am. Chem. Soc. 117, 3340-3346.
Laemmli, U. K. (1970) Nature, 277, 680-685.
Kim, H., Kim, S. G., Lee, M. Y., and Novak, R. F. (1992) Biochem. Biophys. Res. Commun. 186, 846-853.
Kim, H., Putt, D., Reddy, S., Hollenberg, P. F., and Novak, R. F. (1993) J. Pharmacol. Exp. Ther. 267, 927-936.
Hla, T., and Neilson, K., Proc. Natl. Acad. Sci. U.S.A. 89:7384-7388 (1992).
Ye, Y.Z., Strong, M., Huang, Z. and Beckman, J.S. (1996)inMethods in Enzymology (Packer, L. ed) vol. 269, pp. 201-209, Acedemic Press Inc., New York, N. Y.

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