在 MAT 的研究領域上還有很多是未知的，而其在醫藥上也扮演重要角
色。由於過去MAT 活性分析除了放射線同位素法外，一直缺少一個高感度，
又簡便的分析方法。本研究我們建立了一個酵素偶合螢光檢測之液相層析法 來 分 析 MAT 酵 素 活 性 及 酵 素 動 力 學 常 數 。 這 個 方 法 利 用 catechol-O-methyltransferase 將 AdoMet 上的甲基轉移到 esculetin 上形成 scopoletin，接者利用 HPLC 螢光檢測分析這些被甲基化的生成物，進而檢測 出 MAT 的活性及酵素動力學常數。並測得 HL-60 MAT 的 Km = 6.1 ± 0.3 µM，Vmax = 135.4 ± 1.5 nmole AdoMet/mg protein/hr。
這個方法感度高，對 AdoMet 有專一性，分析時干擾少，過程簡便快速，
花費便宜，故此法在MAT 活性分析以及酵素動力學研究上，提供了一個較好
的選擇，將來也可試著應用於臨床診斷與抗憂鬱藥AdoMet 血中濃度監測。
參考文獻
1. Tabor, C.W. and Tabor, H. Methionine adenosyltransferase (S-adenosyl- methionine synthetase) and S-adenosylmethionine decarboxylase. Adv.
Enzymol. Relat. Areas. Mol. Biol., 1984;56:251-282.
2. Kotb, M. and Geller, A.M. Methionine adenosyltransferase: structure and function. Pharmacol. Ther., 1993;59(2):125-143.
3. Mato, J.M., Alvarez, L., Ortiz, P. and Pajares, M.A. S-adenosylmethionine synthesis: molecular mechanisms and clinical implications. Pharmacol. Ther., 1997;73(3):265-280.
4. Mudd, S.H. and Cantoni, G.L. Activation of methionine for transmethylation.
III. The methionine-activating enzyme of Bakers′ yeast. J. Biol.Chem., 1958;231:481-492.
5. Markham, G.D., DeParasis, J. and Gatmaitan, J. The sequence of metK, the structural gene for S-adenosylmethionine synthetase in Escherichia coli. J.
Biol. Chem., 1984;259(23):14505-14507.
6. Thomas, D. and Surdin-Kerjan, Y. SAM1, the structural gene for one of the S-adenosylmethionine synthetases in Saccharomyces cerevisiae. Sequence and expression. J. Biol. Chem., 1987;262(34):16704-16709.
7. Peleman, J., Saito, K., Cottyn, B., Engler, G., Seurinck, J., Van Montagu, M.
and Inze, D. Structure and expression analyses of the S-adenosyl methionine synthetase gene family in Arabidopsis thaliana. Gene, 1989;84(2):359-369.
8. Peleman, J., Boerjan, W., Engler, G., Seurinck, J., Botterman, J., Alliotte, T., Van Montagu, M. and Inze, D. Strong cellular preference in the expression of a housekeeping gene of Arabidopsis thaliana encoding S-adenosylmethionine synthetase. Plant. Cell., 1989;1(1):81-93.
9. Alvarez, L., Corrales, F., Martin-Duce, A. and Mato, J.M. Characterization of a full-length cDNA encoding human liver S-adenosylmethionine synthetase:
tissue-specific gene expression and mRNA levels in hepatopathies. Biochem.
J., 1993;293 (Pt 2):481-486.
10. Horikawa, S. and Tsukada, K. Molecular cloning and developmental expression of a human kidney S-adenosylmethionine synthetase. FEBS Lett., 1992;312(1):37-41.
11. Horikawa, S., Ishikawa, M., Ozasa, H. and Tsukada, K. Isolation of a cDNA encoding the rat liver S-adenosylmethionine synthetase. Eur. J.
Biochem., 1989;184(3):497-501.
12. Horikawa, S., Sasuga, J., Shimizu, K., Ozasa, H. and Tsukada, K. Molecular cloning and nucleotide sequence of cDNA encoding the rat kidney S-adenosylmethionine synthetase. J. Biol. Chem., 1990;265(23):13683-13686.
13. Horikawa, S. and Tsukada, K. Molecular cloning and nucleotide sequence of cDNA encoding the human liver S-adenosylmethionine synthetase.
Biochem. Int., 1991;25(1):81-90.
14. Sakata, S.F., Shelly, L.L., Ruppert, S., Schutz, G. and Chou, J.Y. Cloning and expression of murine S-adenosylmethionine synthetase. J. Biol. Chem., 1993;268(19):13978-13986.
15. De La Rosa, J., Ostrowski, J., Hryniewicz, M.M., Kredich, N.M., Kotb, M., LeGros, H.L. Jr., Valentine, M. and Geller, A.M. Chromosomal localization and catalytic properties of the recombinant alpha subunit of human lymphocyte methionine adenosyltransferase. J. Biol. Chem., 1995;270(37):21860-21868.
16. Kotb, M., Mudd, S.H., Mato, J.M., Geller, A.M., Kredich, N.M., Chou, J.Y.
and Cantoni, G.I. Consensus nomenclature for the mammalian methionine adenosyltransferase genes and gene products. Trends Genet., 1997;13(2):51-52.
17. van Faassen, H. and Berger, R. Separation of different forms of S-adenosylmethionine synthetase by fast protein liquid chromatography. J.
Biochem. Biophys. Methods., 1990 Mar;20(3):189-194.
18. Gil, B., Casado, M., Pajares, M.A., Bosca, L., Mato, J.M., Martin-Sanz, P. and Alvarez, L. Differential expression pattern of S-adenosylmethionine synthetase isoenzymes during rat liver development. Hepatology, 1996 Oct;24(4):876-881.
19. Alvarez, L., Mingorance, J., Pajares, M.A. and Mato, J.M. Expression of rat liver S-adenosylmethionine synthetase in Escherichia coli. results in two active oligomeric forms. Biochem. J., 1994;301 ( Pt 2):557-561.
20. Cabrero, C., Puerta, J. and Alemany, S. Purification and comparison of two forms of S-adenosyl-L-methionine synthetase from rat liver. Eur. J. Biochem., 1987;170(1-2):299-304.
21. Takusagawa, F., Kamitori, S. and Markham, G.D. Structure and function of S-adenosylmethionine synthetase: crystal structures of S-adenosylmethionine synthetase with ADP, BrADP, and PPi at 2.8 angstroms resolution.
Biochemistry, 1996;35(8):2586-2596.
22. Mingorance, J., Alvarez, L., Pajares, M.A. and Mato, J.M. Recombinant rat
liver S-adenosyl-L-methionine synthetase tetramers and dimers are in equilibrium. Int. J. Biochem. Cell Biol., 1997;29(3):485-491.
23. Hoffman, J.L. Fractionation of methionine adenosyltransferase isozymes (rat liver). Methods Enzymol., 1983;94:223-228.
24. Sullivan, D.M. and Hoffman, J.L. Fractionation and kinetic properties of rat liver and kidney methionine adenosyltransferase isozymes. Biochemistry, 1983;22(7):1636-1641.
25. Suma, Y., Shimizu, K. and Tsukada, K. Isozymes of S-adenosylmethionine synthetase from rat liver: isolation and characterization. J. Biochem. (Tokyo), 1986;100(1):67-75.
26. Chamberlin, M.E., Ubagai, T., Pao, V.Y., Pearlstein, R.A. and Yang Chou, J.
Structural requirements for catalysis and dimerization of human methionine adenosyltransferase I/III. Arch. Biochem. Biophys., 2000;373(1):56-62.
27. Kotb, M. and Kredich, N.M. S-Adenosylmethionine synthetase from human lymphocytes. Purification and characterization. J. Biol. Chem., 1985;260(7):3923-3930.
28. Halim, A.B., LeGros, L., Geller, A. and Kotb, M. Expression and functional interaction of the catalytic and regulatory subunits of human methionine
adenosyltransferase in mammalian cells. J. Biol. Chem., 1999;274(42):29720-29725.
29. Wabiko, H., Ochi, K., Nguyen, D.M., Allen, E.R. and Freese, E. Genetic mapping and physiological consequences of metE mutations of Bacillus subtilis. J. Bacteriol., 1988;170(6):2705-2710.
30. Geller, A.M., Legros, H.L., Wherry, K. and Kotb, M.Y. Inhibition of methionine adenosyltransferase by the polyamines. Arch. Biochem. Biophys., 1997;345(1):97-102.
31. Latasa, M.U., Boukaba, A., Garcia-Trevijano, E.R., Torres, L., Rodriguez, J.L., Caballeria, J., Lu, S.C., Lopez-Rodas, G., Franco, L., Mato, J.M. and Avila, M.A. Hepatocyte growth factor induces MAT2A expression and histone acetylation in rat hepatocytes: role in liver regeneration. FASEB J., 2001;15(7):1248-1250.
32. Torre, L., Lopez-Rodas, G., Latasa, M.U., Carretero, M.V., Boukaba, A., Rodriguez, J.L., Franco, L., Mato, J.M. and Avila, M.A. DNA methylation and histone acetylation of rat methionine adenosyltransferase 1A and 2A genes is tissue-specific. Int. J. Biochem. Cell Biol., 2000;32(4):397-404.
33. Gil, B., Pajares, M.A., Mato, J.M. and Alvarez, L. Glucocorticoid regulation
of hepatic S-adenosylmethionine synthetase gene expression. Endocrinology, 1997;138(3):1251-1258.
34. Castro, C., Ruiz, F.A., Perez-Mato, I., Sanchez del Pino, M.M., LeGros, L., Geller, A.M., Kotb, M., Corrales, F.J. and Mato, J.M. Creation of a functional S-nitrosylation site in vitro by single point mutation. FEBS Lett., 1999;459(3):319-322.
35. Perez-Mato, I., Castro, C., Ruiz, F.A., Corrales, F.J. and Mato, J.M.
Methionine adenosyltransferase S-nitrosylation is regulated by the basic and acidic amino acids surrounding the target thiol. J. Biol. Chem., 1999;274(24):17075-17079.
36. Ruiz, F., Corrales, F.J., Miqueo, C. and Mato, J.M. Nitric oxide inactivates rat hepatic methionine adenosyltransferase In vivo by S-nitrosylation.
Hepatology, 1998;28(4):1051-1057.
37. Corrales, F.J., Ruiz, F. and Mato, J.M. In vivo regulation by glutathione of methionine adenosyltransferase S-nitrosylation in rat liver. J. Hepatol., 1999;31(5):887-894.
38. Gomes Trolin, C., Ekblom, J. and Oreland, L. Regulation of methionine adenosyltransferase catalytic activity and messenger RNA in SH-SY5Y
human neuroblastoma cells. Biochem. Pharmacol., 1998;55(5):567-571.
39. Fu, Z., Hu, Y., Markham, G.D. and Takusagawa, F. Flexible loop in the structure of S-adenosylmethionine synthetase crystallized in the tetragonal modification. J. Biomol. Struct. Dyn., 1996;13(5):727-739.
40. Cai, J., Sun, W.M., Hwang, J.J., Stain, S.C. and Lu, S.C. Changes in S-adenosylmethionine synthetase in human liver cancer: molecular characterization and significance. Hepatology, 1996;24(5):1090-1097.
41. Chiang, P.K., Gordon, R.K., Tal, J., Zeng, G.C., Doctor, B.P., Pardhasaradhi, K. and McCann, P.P. S-adenosylmethionine and methylation. FASEB J., 1996;10:471-480.
42. Tajima, S. and Suetake, I. Regulation and function of DNA methylation in vertebrates. Biochem. (Tokyo), 1998;123(6):993-999.
43. Schmutte, C. and Jones, P.A. Involvement of DNA methylation in human carcinogenesis. Biol. Chem., 1998;379(4-5):377-388.
44. Zingg, J.M. and Jones, P.A. Genetic and epigenetic aspects of DNA methylation on genome expression, evolution, mutation and carcinogenesis.
Carcinogenesis, 1997;18(5):869-882.
45. Momparler, R.L., Cote, S. and Eliopoulos, N. Pharmacological approach for
optimization of the dose schedule of 5-aza-2′-deoxycytidine (Decitabine) for the therapy of leukemia. Leukemia, 1997;11(2):175-180.
46. Tutt Landolfi, M.M., Scollay, R. and Parnes, J.R. Specific demethylation of the CD4 gene during CD4 T lymphocyte differentiation. Mol. Immunol., 1997;34(1):53-61.
47. Bender Christina, M., Zingg Jean-Marc and Jones Peter, A. DNA methylation as a target for drug design. Pharmaceutical Research, 1998;15(2):175-187.
48. Potter, V.R., Phenotypic diversity in experimental hepatoma concept of partially blocked ontogeny. Br. J. Cancer., 1978;38:1.
49. Liau, M.C., Chang, C.F. and Giovanella, B.C. Demonstration of an altered S-adenosylmethionine synthetase in human malignant tumors xenografted into athymic nude mice. J. Natl. Cancer Inst., 1980;64:1071.
50. Liau, M.C., Chang, C.F., Saunders, G.F. and Tsai, Y.H.
S-adenosylhomocysteine hydrolases as the primary target enzymes in androgen regulation of methylation complexes. Arch. Biochem. Biophys., 1981;208:261.
51. Liau, M.C. and Burzynski, S.R. Altered methylation complex isozymes as
selective targets for cancer chemotherapy. Drugs Exp. Clin. Res., 1986;12 Suppl 1:77-86.
52. Liau, M.C., Hunt, M.E. and Hurlbert, R.B. Role of ribosomal RNA methylases in the regulation of ribosome production. Biochem., 1976, 15:3158.
53. Hanada, M., Delia, D., Aiello, A., Stadtmauer, E. and Reed, J.C. bcl-2 gene hypomethylation and high-level expression in B-cell chronic lymphocytic leukemia. Blood, 1993;82(6):1820-1828.
54. Crossen, P.E. and Morrison, M.J. Hypermethylation of the M27beta (DXS255) locus in chronic B-cell leukaemia. Br. J. Haematol., 1998;100(1):191-193.
55. Iravani, M., Dhat, R. and Price, C.M. Methylation of the multi tumor suppressor gene-2 (MTS2, CDKN1, p15INK4B) in childhood acute lymphoblastic leukemia. Oncogene, 1997;15(21):2609-2614.
56. Eguchi, K., Kanai, Y., Kobayashi, K. and Hirohashi, S. DNA hypermethylation at the D17S5 locus in non-small cell lung cancers: its association with smoking history. Cancer Res., 1997;57(21):4913-4915.
57. Wicki, R., Franz, C., Scholl, F.A., Heizmann, C.W. and Schafer, B.W.
Repression of the candidate tumor suppressor gene S100A2 in breast cancer is
mediated by site-specific hypermethylation. Cell Calcium, 1997;22(4):243-254.
58. Martin, V., Ribieras, S., Song-Wang, X., Rio, M.C. and Dante, R. Genomic sequencing indicates a correlation between DNA hypomethylation in the 5′
region of the pS2 gene and its expression in human breast cancer cell lines.
Gene, 1995;157(1-2):261-264.
59. Makos, M., Nelkin, B.D., Lerman, M.I., Latif, F., Zbar, B. and Baylin, S.B.
Distinct hypermethylation patterns occur at altered chromosome loci in human lung and colon cancer. Proc. Natl. Acad. Sci. USA, 1992;89(5):1929-1933
60. Huang, Z.Z., Mao, Z., Cai, J. and Lu, S.C. Changes in methionine adenosyltransferase during liver regeneration in the rat. Am. J. Physiol., 1998;275(1 Pt 1):G14-G21.
61. Cai, J., Mao, Z., Hwang, J.J. and Lu, S.C. Differential expression of methionine adenosyltransferase genes influences the rate of growth of human hepatocellularcarcinoma cells. Cancer Res., 1998;58(7):1444-1450.
62. Frago, L.M., Gimenez, A., Rodriguez, E.N. and Varela-Nieto, I. Pattern of methionine adenosyltransferase isoenzyme expression during rat liver
regeneration after partial hepatectomy. FEBS Lett., 1998;426(3):305-308.
63. Newman, E.B., Budman, L.I., Chan, E.C., Greene, R.C., Lin, R.T., Woldringh, C.L. and D′Ari, R. Lack of S-adenosylmethionine results in a cell division defect in Escherichia coli. J. Bacteriol., 1998;180(14):3614-3619.
64. Chamberlin, M.E., Ubagai, T., Mudd, S.H., Levy, H.L. and Chou, J.Y.
Dominant inheritance of isolated hypermethioninemia is associated with a mutation in the human methionine adenosyltransferase 1A gene. Am. J. Hum.
Genet., 1997;60(3):540-546.
65. Nagao, M. and Oyanagi, K. Genetic analysis of isolated persistent hypermethioninemia with dominant inheritance. Acta. Paediatr. Jpn., 1997;39(5):601-606.
66. Mato, I.P., Sanchez del Pino, M.M., Chamberlin, M.E., Mudd, S.H., Mato, J.M. and Corrales, F.J. Biochemical basis for the dominant inheritance of hypermethioninemia associated with the R264H mutation of the MAT1A gene.
A monomeric methionine adenosyltransferase with tripolyphosphatase activity.
J. Biol. Chem., 2001;276(17):13803-13809.
67. Surtees, R., Leonard, J. and Austin, S. Association of demyelination with deficiency of cerebrospinal-fluid S-adenosylmethionine in inborn errors of
methyl-transfer pathway. Lancet., 1991;338 (8782-8783):1550-1554.
68. Chamberlin, M.E., Ubagai, T., Mudd, S.H., Wilson, W.G., Leonard, J.V. and Chou, JY. Demyelination of the brain is associated with methionine adenosyltransferase I/III deficiency. J. Clin. Invest., 1996;98(4):1021-1027.
69. Snyder, S.W., Egorin, M.J. and Callery, P.S. Induction of human leukemia cell differentiation by regiospecifically acetylated spermidines. Biochem. Biophys.
Res. Commun., 1991;180(2):591-596.
70. Bjersing, J.L., Brorsson, A. and Heby, O. Increased expression of c-jun, but not retinoic acid receptor beta, is associated with F9 teratocarcinoma stem cell differentiation induced by polyamine depletion. J. Cell. Biochem., 1997;67(3):378-385.
71. Luk, G.D., Civin, C.I., Weissman, R.M. and Baylin, S.B. Ornithine decarboxylase: essential in proliferation but not differentiation of human promyelocytic leukemia cells. Science, 1982;216(4541):75-77.
72. Sugiura, M., Shafman, T., Mitchell, T., Griffin, J. and Kufe, D. Involvement of spermidine in proliferation and differentiation of human promyelocytic leukemia cells. Blood, 1984;63(5):1153-1158.
73. Riscoe, M.K., Schwamborn, J., Ferro, A.J., Olson, K.D. and Fitchen, J.H.
Inhibition of growth but not differentiation of normal and leukemic myeloid cells by methylthioadenosine. Cancer Res., 1987;47(14):3830-3834.
74. Sydow, K. and Boger, R.H. Homocysteine, endothelial dysfunction and cardiovascular risk: pathomechanisms and therapeutic options. Z. Kardiol., 2001;90(1):1-11.
75. Kaletha, K., Chodorowski, Z., Sein-Anand, J., Gazda, M. and Nagel- Starczynowska, G. Homocysteine as a risk factor for atherosclerosis. Przegl.
Lek., 2000;57(10):591-595.
76. Lu, S.C. S-Adenosylmethionine. The International Journal of Biochemistry &
Cell Biology., 2000;32(4):391-395.
77. Bottiglieri, T., and Hyland, K. S-adenosylmethionine levels in psychiatric and neurological disorders: a review. Acta. Neurol. Scand., 1994;154:19-26.
78. Fava, M., Giannelli, A., Rapisarda, V., Patralia, A. and Guaraldi, G.P.
Rapidity of onset of the antidepressant effect of parenteral S-adenosyl-L-methionine. Psychiatry Res., 1995;56(3):295-297.
79. Bottiglieri, T., Hyland, K. and Reynolds, E.H. The clinical potential of ademetionine(S-adenosylmethionine) in neurological disorders. Drugs, 1994;48(2):137-152.
80. Bressa, G.M. S-adenosyl-l-methionine (SAMe) as antidepressant:
meta-analysis of clinical studies. Acta. Neurol. Scand. Suppl., 1994;154:7-14.
81. Bottiglieri, T., Godfrey, P., Flynn, T., Carney, M.W., Toone, B.K. and Reynolds, E.H. Cerebrospinal fluid S-adenosylmethionine in depression and dementia: effects of treatment with parenteral and oral S-adenosylmethionine.
J. Neurol .Neurosurg. Psychiatry, 1990;53(12):1096-1098.
82. Schumacher, H.R. Jr. Osteoarthritis: the clinical picture, pathogenesis, and management with studies on a new therapeutic agent, S-adenosylmethionine.
Am. J. Med., 1987;83(5A):1-4.
83. Liau, M.C., Chang, C.F. and Becker, F.F. Alteration of S-adenosylmethionine synthetasesduring chemical hepatocarcinogenesis and in resulting carcinomas.
Cancer Res., 1979;39(6 Pt 1):2113-2119.
84. Jennifer, Lagendijk, Johan, B., Ubbink, W.J. and Hayward, Vermaak Quantification of erythrocyte S-adenosyl-L-methionine levels and its application in enzyme studies. J. Chromatogr., 1992;576:95-101 Biomedical Applications.
85. Carolyn K. Wise, Craig A. Gooney, Syed F. Ali and Lionel A. Poirier Measuring S-adenosylmethionine in whole blood, red blood cells and cultured
cells using-a fast preparation method and high performance liquid chromatography. J. Chromatogr. B, 1997;696:145-152.
86. Capdevila, A. and Wagner, C. Measurement of plasma S-adenosylmethionine and S-adenosylhomocysteine as their fluorescent isoindoles. Anal. Biochem., 1998;264(2):180-184.
87. She, Q.B., Nagao, I., Hayakawa, T. and Tsuge, H. A simple HPLC method for the determination of S-adenosylmethionine and S-adenosyl- homocysteine in rat tissues: the effect of vitamin B6 deficiency on these concentrations in rat liver. Biochem. Biophys. Res. Commun., 1994;205(3):1748-1754
88. Hamedani, M.P., Valko, K., Qi, X., Welham, K.J. and Gibbons, W.A.
Two-dimensional high-performance liquid chromatographic method for assaying S-adenosyl-L-methionine and its related metabolites in tissues. J Chromatogr., 1993;619(2):191-198.
89. Valko, K., Hamedani, M.P., Ascah, T.L. and Gibbons, W.A. A comparative study of the reversed-phase HPLC retention behaviour of S-adenosyl-L-methionine and its related metabolites on Hypersil ODS and Supelcosil LC-ABZ stationary phases. J. Pharm. Biomed. Anal., 1993;11(4-5):361-366.
90. Bottiglieri, T. Isocratic high performance liquid chromatographic analysis of S-adenosylmethionine and S-adenosylhomocysteine in animal tissues: the effect of exposure to nitrous oxide. Biomed. Chromatogr., 1990;4(6):239-241.
91. Wagner, J., Claverie, N. and Danzin, C. A rapid high-performance liquid chromatographic procedure for the simultaneous determination of methionine, ethionine, S-adenosylmethionine, S-adenosylethionine, and the natural polyamines in rat tissues. Anal. Biochem., 1984;140(1):108-116.
92. Wagner, J., Danzin, C., Huot-Olivier, S., Claverie, N. and Palfreyman, M.G.
High-performance liquid chromatographic analysis of S-adenosylmethionine and its metabolites in rat tissues: interrelationship with changes in biogenic catechol levels following treatment with L-dopa. J. Chromatogr., 1984;290:247-262.
93. Gutheil, W.G., Stefanova, M.E. and Nicholas, R.A. Fluorescent coupled enzyme assays for D-alanine: application to penicillin-binding protein and vancomycin activity assays. Anal. Biochem., 2000;287(2):196-202.
94. Yasmineh, W.G., Chung, M.Y. and Caspers, JI. Determination of serum catalase activity on a centrifugal analyzer by an NADP/NADPH coupled enzyme reaction system. Clin. Biochem., 1992;25(1):21-27.
95. Alon, R., Bayer, E.A. and Wilchek, M. Related Articles A coupled enzyme assay for measurement of sialidase activity. J. Biochem. Biophys. Methods., 1991;22(1):23-33.
96. Van Lente, F. and Pepoy, M. Coupled-enzyme determination of catalase activity in erythrocytes. Clin. Chem., 1990;36(7):1339-1343.
97. Hirashima, Y., Farooqui, A.A. and Horrocks, L.A. Related Articles Fluorimetric coupled enzyme assay for lysoplasmalogenase activity in liver.
Biochem. J., 1989;260(2):605-608.
98. Tsai, S.F., Bishop, D.F. and Desnick, R.J. Coupled-enzyme and direct assays for uroporphyrinogen III synthase activity in human erythrocytes and cultured lymphoblasts. Enzymatic diagnosis of heterozygotes and homozygotes with congenital erythropoietic porphyria. Anal. Biochem., 1987;166(1):120-133.
99. Teruel, J.A., Tudela, J., Fernandez-Belda, F., Garcia-Carmona, F., Garcia-Canovas, F. and Gomez-Fernandez, J.C. A kinetic study of the irreversible inhibition of an enzyme measured in the presence of coupled enzymes. Fluorescein isothiocyanate as inhibitor of the adenosinetriphosphatase activity from sarcoplasmic reticulum. Biochim.
Biophys. Acta., 1986;869(1):8-15.
100. Ozer, N. A new enzyme-coupled spectrophotometric method for the determination of arginase activity. Biochem. Med., 1985;33(3):367-371.
101. Wilkinson, A.P., Rhodes, M.J. and Fenwick, G.R. Determination of
myrosinase (thioglucoside glucohydrolase) activity by a spectrophotometric coupled enzyme assay. Anal. Biochem., 1984;139(2):284-291.
102. Giampietro, P.F. and Desnick, R.J. Determination of delta-aminolevulinate dehydratase activity by a specific fluorometric coupled-enzyme assay. Anal.
Biochem., 1983;131(1):83-92.
103. Nohta, H., Noma, S. and Ohkura, Y. Assay for Catechol-O- methyltransferase in erythrocytes using a new fluorogenic substrate, 2-(3,4-Dihydroxyphenyl) naphtho[1,2-d]thiazole. J. Chromatogr., 1984; 304:93-100.
104. Nohta, H., Noma, S. and Ohkura, Y. High-performance liquid
chromatographic assay for Catechol-O-methyltransferase in human and rat erythrocyte membrane and soluble fractions, and rat tissues. Chem. Pharm.
bull., 1986;34(11):4687-4693.
105. Muller-Enoch, D., Seidl, E. and Thomas, H. [6.7-Dihydroxycoumarin
(Aesculetin) as a substrate for catechol-O-methyltransferase (author′s transl)].
Z. Naturforsch. [C], 1976;31(5-6):280-284.
106. Thomas, H., Veser, J. and Muller-Enoch, D. [A sensitive fluorometric
determination of catechol methyltransferase activity (author′s transl)]. Hoppe.
Seylers. Z. Physiol. Chem., 1976;357(10):1347-1351.
107. Hoffman, R.M. Methioninase: a therapeutic for diseases related to altered methionine metabolism and transmethylation: cancer, heart disease, obesity, aging, and Parkinson′s disease. Hum. Cell, 1997;10(1):69-80.
108. Cheng, H., Gomes-Trolin, C., Aquilonius, S.M., Steinberg, A., Lofberg, C., Ekblom, J. and Oreland, L. Levels of L-methionine S-adenosyltransferase activity in erythrocytes and concentrations of S-adenosylmethionine and S-adenosylhomocysteine in whole blood of patients with Parkinson′s disease.
Exp. Neurol., 1997;145(2 Pt 1):580-585.
109. Bradford, M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal.
Biochem., 1976;72,248-254.
110. Trolin, C.G., Lofberg, C., Trolin, G. and Oreland, L. Brain ATP:L-methionine S-adenosyltransferase (MAT), S-adenosylmethionine (AdoMet) and
S-adenosylhomocysteine (SAH): regional distribution and age-related changes.
Eur. Neuropsychopharmacol, 1994;4(4):469-477.
111. Ekegren, T., Aquilonius, S.M. and Gomes-Trolin, C. A comparative study of methionine adenosyltransferase activity and regional distribution in
mammalian spinal cord. Biochem. Pharmacol., 2000;60(3):441-445.
112. Tu, Y.S., Huang, L.J., Hsu, M.H., Wang, S.H., Lai, J.M., Yeh, C.C., Liau, M.C.
and Lung, F.D. Synthesis of hemoregulatory peptide (HP) derivatives as inhibitors of the interaction between methionine adenosyl transferase and methionine. The Chin. Pharm. J., 2000;52:139-149.
113. Gallagher, R., Collins, S., Trujillo, J., McCredie, K., Ahearn, M., Tsai, S., Metzgar, R., Aulakh, G., Ting, R., Ruscetti, F., and Gallo, R. Characterization of the continuous, differentiating myeloid cell line (HL-60) from a patient with acute promyelocytic leukemia. Blood, 1979; 54:713-733.
114. Liau, M. C., Lin, G. W., and Hurlbert, R. B. Partial purification and
characterization of tumor and liver S-adenosylmethionine synthetases. Cancer Res., 1977;37:427-435.
115. Hoffman, J. L. Chromatographic analysis of the chiral and covalent instability of S-adenosyl-L-methionine. Biochemistry, 1986;25:4444-4449.
116. Revelle, L. K., d′Avignon, D. A., Reepmeyer, J.C. and Zerfing, R. C.
Stability-indicating proton nuclear magnetic resonance spectroscopic method for determination of S-adenosyl-L-methionine in tablets. J. AOAC Int.,
1995;78:353-358.
中國醫藥大學 藥物化學研究所 博士論文
(二)
指 導 教 授 ： 郭盛助 教 授 指 導 教 授 ： 陳勝智 教 授
HotLig: 以分子表面導向之方法評估蛋白質-配體之交互作用
HotLig: A molecular surface-directed approach to scoring protein-ligand interactions
研究生： 王 勝 弘
Wang, Sheng-Hung
中 華 民 國 95 年 6 月
目錄
收集用於分析蛋白質-配體交互作用之統計組... 33
圖目錄
Scheme 1. HotLig 之設計與發展... 25 圖 1. Type of Lennard-Jones 12-6 dispersion-repulsion energy function ... 7 圖 2. 疏水效應示意圖... 19 圖 3. Solvent-accessible surface... 23 圖 4. Connolly surface... 24 圖 5. 網格座標區域設定 PscanMS 掃描範圍 ... 27 圖 6. 移動掃描格並選取蛋白質原子... 28 圖 7. 掃描球在掃描格內進行掃描... 28 圖 8. 掃描點之紀錄... 29 圖 9. Binding pocket 掃描結果... 30 圖 10. 組成Connolly surface 之三種表面描述 ... 31 圖 11. 建構Connolly surface 之三種基本幾何圖形 ... 31 圖 12. 十種已知的HSV-1 thymidine kinase 配體結構... 43 圖 13. 統計組 600 個複合結構中配體結構特性之分布... 47 圖 14. 氫鍵作用力之統計結果與勢能曲線... 49 圖 15. 離子作用力之統計結果與勢能曲線... 50
圖 16. 金屬配位共價鍵作用力之統計結果與勢能曲線... 51 圖 17. 疏水效應非極性區域之統計結果與勢能曲線... 52 圖 18. 訓練組 214 個複合結構中配體結構特性之分布... 56 圖 19. HotLig 之蛋白質-配體親和力預測值與實驗數據值相關圖 ... 63 圖 20. HSV-1 thymidine kinase (TK) 之虛擬篩選測試... 67
表目錄
表 1. Novel ligands recently identified through structure-based VS. ... 2
表 2. 現行各種評分方程式蛋白質-配體結合模式預測成功率之
比較... 17 表 3. 現行評分方程式在不同性質subsets 結合模式預測成功率之
差異... 18 表 4. Statistics set ... 34 表 5. Training set... 39 表 6. The optimized parameters and the best success rate of
predicting binding modes of training sets... 57 表 7. HotLig 與其他評分方程式對 Wang 100 dataset 預測結合模
predicting binding modes of training sets... 57 表 7. HotLig 與其他評分方程式對 Wang 100 dataset 預測結合模