ISOLATION/SYNTHESES, CELLULAR ABSORPTION, AND MOLECULAR FUNCTIONS OF BIO-ACTIVE PHYTOCHEMICALS

Jae B. Park, Ph.D. BHNRC, ARS, USDA

Numerous epidemiological studies have correlated human consumption of diets rich in fruits and vegetables containing high levels of phytochemicals to lower risk for specific chronic diseases such as diabetes, cancer and cardiovascular diseases. Phytochemicals are widely distributed in plant-derived foods of the typical U.S. diet. These compounds have been reported to contribute to numerous purported beneficial health effects. However, understanding the specific biological mechanisms involved, relative to human diseases, presents challenging and researchable taskes. Without adequate definitive information on cellular absorption and molecular functions of phytochemicals, it is not possible to accurately assess their beneficial effects on human health. Moreover, optimal but safe intakes of phytochemicals have yet to be determined; these data must be generated before any recommendations of phytochemicals can be made. Therefore, a research project has been proposed to gain information on the cellular absorption and molecular functions of various phytochemicals, which might lead us to a better understanding of cellular and molecular mechanisms and to elucidating their beneficial effects.

The proposed research is particularly focused on phytochemicals from several plants including wolfberry, safflower, coco and other plants with potential health effects.

Objectives

1. Purification and/or Syntheses of Bio- Active Phytochemicals:
To purify and/or synthesize bio-active phytochemicals.
2. Bio-availability of Phytochemicals:
To delineate the mechanisms of phytochemical absorption and determine bio-availability of phytochemicals.
3. Cellular and Molecular Actions of Phytochemicals:
To determine cellular and molecular mechanisms related to human diseases such as obesity, diabetes, heart diseases, cancer and aging.
4. Construction of Inter-related Plant, Phytochemical and Bio-activity Databases related to Human Chronic Diseases:
To build inter-related plant, phytochemical and activity databases related to human diseases such as inflammation, obesity, diabetes, heart diseases, cancer and aging.

1. Park, J.B., Chae, H.Z. & Kim, Y.S. (1986) "Purification and characterization glyoxyate synthase in Acinetobacter calcoaceticus." K. Biochemistry Journal 19, 235-241.

2. Conover, R.C., Park, J.B., Adams, M.W.W., & Johnson, M.K. (1990) "The formation and properties of a NiFe3S4 cluster in Pyrococcus furiosus ferredoxin." J. Amer. Chem. Society 112, 4562-4564

3. Conover, R.C., Kowal, A.T., Fu, W., Park, J.B., Aono, S., Adams, M.W.W., & Johnson, M.K. (1990) "Spectroscopic Characterization of the Novel Iron-Sulfur Cluster in Pyrococcus furiosus Ferredoxin." J. Biol. Chem. 265, 8533-8541.

4. Park, J.B., Fan, C., Hoffmans, B.M., & Adams, M.W.W. (1991) "Potentiometric and electron nuclear double resonance properties of two spin forms of the [4Fe-4S]+ Cluster in the Novel Ferredoxin from Hyperthermophilic Archaebacterium Pyrococcus furiosus." J. Biol. Chem. 266, 19351-19356.

5. Conover, R.C., Park, J.B., Adams, M.W.W. & Johnson, M.K. (1991) "Exogenous ligand binding to the [Fe4S4] cluster in Pyrococcus furiosus ferredoxin" J. Amer. Chem. Society, 113, 2799-2800.

6. Blake, P.R., Park, J.B., Bryant, F.O., Aono, S., Magnuson, J.K., Eccleston, E., Howard, J.B., Summers, M.F., & Adams, M.W.W. (1991) "Determinants of Protein Hyperthermostability: Purification and Amino Acid Sequence of Rubredoxin from the Hyperthermophilic Archaebacterium Pyrococcus furiosus and Secondary Structure of the Zinc Adduct by NMR." Biochemistry, 30, 10885-10895.

7. Robb, F.T., Park, J.B., & Adams, M.W.W. (1992) "Characterization of an extremely thermostable glutamate dehydrogenase: a key enzyme in the primary metabolism of the hyperthermophilic archaebacterium, Pyrococcus furiosus." Biochimica et Biophysica Acta 1120, 267-272.

8. Robb, F.T., Masuchi, Y., Park, J.B. & M.W.W. Adams (1992) "Biocatalysis at extreme temperatures." J. American Chemical Society, 74-86.

9. Klump, H., Ruggiero, J.D., Kessel, M., Park, J.B., Adams, M.W.W., & Robb, F.T. (1992) "Glutamate Dehydrogenase from the Hyperthermophile Pyrococcus furiosus." J. Biol. Chem. 267, 22681-22685.

10. George, S.J., Chen, C.T., Park, J.B. Park & Adams, M.W.W. (1992) J. Amer. Chem. Society 114, 4426-4427.

11. Day, M.W., Hsu, B.T., Joshua-Tor, L., Park, J.B., Zhou, Z.H., Adams, M.W.W., & Rees, D.C. (1992) "X-ray crystal structures of the oxidized and reduced forms of the rubredoxin from the marine hyperthermophilic archaebacterium Pyrococcus furiosus" Protein Science 1, 1494-1507.

12. Blake, P.R., Day, M.W., Hsu, B.T., Joshua-Tor, L., Park, J.B., Hare, D.R., Adams, M.W.W., Rees, D.C., & Summers, M.F. (1992) "Comparison of the X-ray structure of native rubredoxin from Pyrococcus furiosus with the NMR structure of the zinc-substituted protein" Protein Science, 1522-1525.

13. Blake, P.R., Park, J.B., Zhou, Z.H., Hare, D.R., Adams, M.W.W., & Summers, M.F. (1992) "Solution-state structure by NMR of zinc-substituted rubredoxin from the marine hyperthermophilic archaebacterium Pyrococcus furiosus" Protein Science 1, 1508-1521.

14. Blake, P.R., Park, J.B., Adams, M.W.W. & Bax, A. (1992) "Quantitative measurement of small through-hydrogen-bond and thruogh-space 1H-113Cd and 1H-199Hg J-couplings in metal-substituted rubredoxin from Pyrococcus furiosus" J. Biomol. NMR 2, 527-533.

15. Blake, P.R., Park, J.B. & M.F. Summers (1992) "Metal-substituted rebredoxin from Pyrococcus furiosus" J. Amer. Chem. Society 114, 4931-4933.

16. Adams, M.W.W., Park, J.B. & Kelly, R.M. (1992) "Biocatalysis at extreme temperatures" J. Amer. Chem. Society, 4-23.

17. Srivastava, K.K.P., Surerus, K.K., Conover, R.C., Johnson, M.K., Park, J.B., Adams, M.W.W., & Munck, E. (1993) "Mossbauer Study of ZnFe3S4 Clusters in Pyrococcus furiosus Ferredoxin" Inorg. Chem., 32, 927-936.

18. Schicho, R.N., Snowden, L.J., Mukund, S., Park, J.B., Adams, M.W.W., & Kelly, R.M. (1993) "Influence of tungsten on metabolic patterns in Pyrococcus furiosus, a hyperthermophilic archaeon" Arch. Microbiol., 159, 380-385.

19. Huang, Y.H., Moura, I., Moura, J.J.G., LeGall, J., Park, J.B., Adams, M.W.W., & Johnson, M.K. (1993) "Resonance Raman Studies of Nickel Tetrathiolates and Nickel-Substituted Rubredoxins and Desulforedoxin" Inorg. Chem., 32, 406-412.

20. Park, J.B., Ashizawa, K., Parkison, C., & Cheng, S. (1993) "One-step immunoaffinity purification of human B1 thyroid hormone receptor with DNA and hormone binding activity" J. Biochem. & Biophys. Methods. , 60, 127-129.

20. Levine, M., Dhariwal, K., Welch, R., Wang, Y., & Park, J.B. (1995) "Determination of optimal vitamin C requirements in humans.". Amer J. Clin. Nutr. 62 (suppl), 1347S-1356S.

22. Welch, R., Wang, Y., Crossman, A., Park, J.B., Kirk, K. L. & Levine, M. (1995) "Accumulation of Vitamin C and Its Oxidized Metabolite Dehydroascorbic Acid Occurs by Separate Mechanisms." J. Biol. Chem. 270, 12584-12592.

23. Levine, M., Cantilena, C.C., Wang, Y., Welch, R.W., Washko, P.W., Dariwal, K.R., & Park, J.B., Graumlich, J.P., King, J., and Cantilena, L.R. (1996) "Vitamin C pharmacokinetics in healthy volunteers: evidence for a Recommended Dietary Allowance." Proc. Natl. Acad. Sci. U.S.A. 93, 3704-3709.

24. Park, J.B., & Levine, M., (1996) "Purification, Cloning, and Expression of Dehydroascorbic Acid Reduction Activity From Human Neutrophils: Identification As Glutaredoxin." Biochem J. 315, 931-938.

25. Park, J.B., & Levine, M., (1997) "The Human Glutaredoxin: Determination of Its Organization, Transcription Start Point, and Promoter Analysis." Gene 197, 189-193

26. Levine, M., Rumsey, S. C., Wang, Y., Park, J. B., Kwon, O., & Amano, N. (1997) "In situ kinetics: an approach to recommended intake of vitamin C." Method of Enzymology, 281: 425-437.

27. Levine M, Rumsey, S.C., Daruwala, R., Park, J.B. & Wang Y. (1999) "Criteria and Recommendations for Vitamin C intake." JAMA, 281, 1415-1422.

28. Levine, M., Darwuala, R., Park, J. B., Rumsey, S. C., & Wang, Y. (1998) A Does vitamin C have a pro-oxidant effect? Nature 395: 231-232.

29. Park, J. B. (1999) "Flavonoids are potential inhibitors of glucose uptake in U937 cells." BBRC, 260, 568-574.

30. Park, J. B. & Levine, M. (2000) "Characterization of Human Ribonucleotide Reductase M2 Subunit." BBRC, 267, 651-657.

31. Park, J. B. & Levine M (2000) "Intracellular Accumulation of Ascorbic Acid is inhibited by Flavonoids via Blocking both dehydroascorbic acid and ascorbic acid uptake in the transformed cells." J. Nutri., 130, 1297-1302.

32. Park, J. B. (2001)"Inhibition of Glucose and Dehydroascorbic Acid Uptake by Resveratrol in Human Transfomed Myeolocytic Cells." J. Nat. Prod. 64, 381-384.

33. Park, J.B. (2001) "A New Reporter Vector Containing both Photnus pyralis and Renilla reniformis Luciferase Genes." Analytical Biochemistry 291, 162-166.

34. Park, J. B. (2001) " Reduction of Dehydroascorbic acid by Homocysteine." BBA 1525, 173-179.

35. Park, J.B. & Schoene, N. (2002) "Synthesis and characterization of N-coumaroyltyramine as a potent phytochemical which arrests human transformed cells via inhibiting protein tyrosine kinases." BBRC 292, 1104-1110.

36. Song, J., Kwon, O., Chen, S., Daruwala, R., Eck, P., Park, J.B. & Levine M.(2002) "Flavonoid inhibition of sodium-dependent vitamin C transporter 1(SVCT1) and glucose transporter isoform 2 (Glut2), intestinal transporters for vitamin C and glucose." JBC 277, 15252-15260

37. Park, J.B., Schoene, N. (2003) "N-Caffeoyltyramine arrests growth of U937 and Jurkat cells by inhibiting protein tyrosine phosphorylation and inducing caspase-3." Cancer Lett. 202, 161-171.

38. Park, J. B. and Mark, L (2005) Cloning, sequencing, and characterization of alternatively spliced glutaredoxin 1 cDNA and its genomic gene: chromosomal localization, mrna stability, and origin of pseudogenes. J Biol Chem. 280, 10427-34.

39. Park, J B, (2005) "N-coumaroyldopamine and N-caffeoyldopamine increase cAMP via beta 2-adrenoceptors in myelocytic U937 cells." FASEB J. 19 497-502.

40. Park, J.B. (2005) "Quantitation of clovamide-type phenylpropenoic acid amides in cells and plasma using high-performance liquid chromatography with a coulometric electrochemical detector." J. Agric. Food. Chem. 2005 Oct 19;53(21):8135-40.

41. Park, J.B., Schoene, N. (2006) "Clovamide-type phenylpropenoic acid amides, N-coumaroyldopamine and N-caffeoyldopamine, inhibit platelet-leukocyte interactions via suppressing P-selectin expression." J Pharmacol Exp Ther. 317 813-819.

42. Park, J.B.(2007) "Caffedymine from cocoa has COX inhibitory activity suppressing the expression of a platelet activation marker, P-selectin." J Agric Food Chem. 55, 2171-5.

43. Park, J.B., Chen, P.(2007) "Synthesis of safflomide and its HPLC measurement in mouse plasma after oral administration." J Chromatogr B Analyt Technol Biomed Life Sci. 852, 398-402.

44. Park, J.B. (2008) SEROTOMIDE AND SAFFLOMIDE MODULATE FORSKOLIN-STIMULATED CAMP FORMATION VIA 5-HT1 RECEPTOR. Phytomedicine. 15, 1093-1098.  

45. Park, J.B. (2008) 5-Caffeoylquinic Acid And Caffeic Acid Orally Administrated Suppresses P-Selectin Expression on Mouse Platelets. J Nutr Biochem. 20, 800-805.  

46. Park, J.B. (2009) Isolation and Characterization of N-Feruloyltyramine as the P-Selectin Expression Suppressor from Garlic (Allium sativum). J Agric Food Chem. 57, :8868-8872.  

47. Park, J.B. (2010) Effects of Typheramide and Alfrutamide Found in Allium Species on Cyclooxygenases and Lipoxygenases. J Med Food 14, 226–231.

48. Park, J.B. (2010) Synthesis, Biological Activities and Bioavailability of Moschamine, a Safflomide-type Phenylpropenoic Acid Amide found in Centaurea cyanus. Natural Product Research .

49. Park, J.B. (2011) Identification and quantification of a major anti-oxidant and anti-inflammatory phenolic compound found in basil, lemon thyme, mint, oregano, rosemary, sage, and thyme. International Journal of Food Science and Nutrition.

50. Park, J.B. (2011) Protective Effects of Veskamide, Enferamide, Becatamide, and Oretamide on H2O2-Induced Apoptosis of PC-12 Cells. Phytomedicine.

Books and Journal Articles

Levine, M, Dhariwal, K.R., Wang, Y., Park, J.B. and Welch, R.W. (1994) Ascorbic acid in neutrophils, in Natural Antioxidants in Human Health and Disease, B. Frei, ed, Academic Press. 469-484.

Levine, M., Rumsey, S., Wang, Y., Park, J.B., Kwon, O., Xu, W., and Amano, N. (1996)
Vitamin C in Present Knowledge in Nutrition, 7th edition, Filer, L.J., Ziegler, E.E., editors. Washington D.C. International Life Science Institute.

Levine M., Rumsey, S.C., Wang, Y., Park, J. B., Daruwala, R., and Amano, N. (1998) Recommended vitamin C intake: from molecular mechanisms to clinical application; in Proceedings of the International Symposium on Vitamin C: the state of the art in disease preventation sixty years after the Novel Prize, Paoletti R., Sies H., Bug J., Grossi E., and Poli A. editors. Springer-Verlag, Milan, pages 107-123.

Levine M., Rumsey, S.C., Wang, Y., Park, J. B., and Daruwala, R. (1998) Vitamin C. In Biochemical and Physiological Bases of Human Nutrition, Stipanuk M., ed. W.B. Saunders.

Park, J. B.(2003) Quercetin. In Encyclopedia of Dietary Supplement 1st, Marcel Dekker, New York, NY

Park, J. B.(2010) Quercetin. In Encyclopedia of Dietary Supplement 2nd , Marcel Dekker, New York, NY