BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
권호 표지
DOI QR코드

DOI QR Code

Human Brain Pyridoxal-5'-phosphate Phosphatase: Production and Characterization of Monoclonal Antibodies

  • Kim, Dae-Won (Department of Anatomy, College of Medicine, Hallym University) ;
  • Eum, Won-Sik (Department of Anatomy, College of Medicine, Hallym University) ;
  • Choi, Hee-Soon (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Kim, So-Young (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • An, Jae-Jin (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Lee, Sun-Hwa (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Sohn, Eun-Joung (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Hwang, Seok-Il (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Kwon, Oh-Shin (Department of Biochemistry, Kyungpook National University) ;
  • Kang, Tae-Cheon (Department of Anatomy, College of Medicine, Hallym University) ;
  • Won, Moo-Ho (Department of Anatomy, College of Medicine, Hallym University) ;
  • Cho, Sung-Woo (Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine) ;
  • Lee, Kil-Soo (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Park, Jin-Seu (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Choi, Soo-Young (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University)
  • Published : 2005.11.30
Download PDF
⟨ Previous Next ⟩

Abstract

We cloned and expressed human pyridoxal-5'-phosphate (PLP) phosphatase, the coenzymatically active form of vitamin $B_6$, in Escherichia coli using pET15b vector. Monoclonal antibodies (mAb) were generated against purified human brain PLP phosphatase in mice, and four antibodies recognizing different epitopes were obtained, one of which inhibited PLP phosphatase. The binding affinities of these four mAbs to PLP phosphatase, as determined using biosensor technology, showed that they had similar binding affinities. Using the anti-PLP phosphatase antibodies as probes, we investigated their cross-reactivities in various mammalian and human tissues and cell lines. The immunoreactive bands obtained on Western blots had molecular masses of ca. 33 kDa. Similarly fractionated extracts of several mammalian cell lines all produced a single band of molecular mass 33 kDa. We believe that these PLP phosphatase mAbs could be used as valuable immunodiagnostic reagents for the detection, identification, and characterization of various neurological diseases related to vitamin $B_6$ abnormalities.

Keywords

References

  1. Anderson, B. B., Newmark, P. A., Rawlins, M. and Green, R. (1974) Plasma binding of vitamin $B_6$ compounds. Nature 250, 502-504 https://doi.org/10.1038/250502a0
  2. Bull, H., Murray, P. G., Thomas, A. M. and Nelson, P. N. (2002) Acid phosphatases. Mol. Pathol. 55, 65-72 https://doi.org/10.1136/mp.55.2.65
  3. Buss, D. D., Hamm, M. W., Mehansho, H. and Henderson, L. M. (1980) Transport and metabolism of pyridoxine in the perfused small intestine and the hind limb of the rat. J. Nutr. 110, 1655- 1663 https://doi.org/10.1093/jn/110.8.1655
  4. Choi, E. Y., Park, S. Y., Jang, S. H., Song, M. S., Cho, S. W. and Choi, S. Y. (1995) Production and characterization of monoclonal antibodies to bovine brain succinic semialdehyde reductase. J. Neurochem. 64, 371-377 https://doi.org/10.1046/j.1471-4159.1995.64010371.x
  5. Choi, E. Y., Jang, S. H. and Choi, S. Y. (1996) Human brain GABA transaminase is immunologically distinct from those of other mammalian brain. Neurochem. Int. 28, 597-600 https://doi.org/10.1016/0197-0186(95)00111-5
  6. Eum, W. S., Choi, H. S., Kim, D. W., Jang, S. H., Choi, S. H., Kim, S. Y., Park, J., Kang, J. H., Cho, S. W., Kwon, O. S., Hwang, I. K., Yoo, K. Y., Kang, T. C., Won, M. H. and Choi, S. Y. (2005) Production and characterization of monoclonal antibodies against human ceruloplasmin. J. Biochem. Mol. Biol. 38, 71-76 https://doi.org/10.5483/BMBRep.2005.38.1.071
  7. Fagerstam, L. G., Karlsson, A. F., Karlsson, R., Persson, B. and Ronnberg, I. (1992) Biospecific interaction analysis using surface plasmon resonance detection applied to kinetic, binding site and concentration analysis. J. Chromatogr. 597, 397-410 https://doi.org/10.1016/0021-9673(92)80137-J
  8. Fonda, M. L. (1992) Purification and characterization of vitamin $B_6$-phosphate phosphatase from human erythrocytes. J. Biol. Chem. 267, 15978-15983
  9. Fonda, M. L. and Zhang, Y. (1995) Kinetic mechanism and divalent metal activation of human erythrocyte pyridoxal phosphatase. Arch. Biochem. Biophys. 320, 345-352 https://doi.org/10.1016/0003-9861(95)90018-7
  10. Gao, G. and Fonda, M. L. (1994a) Kinetic analysis and chemical modification of vitamin $B_6$ phosphatase from human erythrocytes. J. Biol. Chem. 269, 7163-7168
  11. Gao, G. and Fonda, M. L. (1994b) Identification of an essential cysteine residue in pyridoxal phosphatase from human erythrocytes. J. Biol. Chem. 269, 8234-8239
  12. Gao, G. and Fonda, M. L. (1994c) Evidence for a phosphoenzyme intermediate formed during catalysis by pyridoxal phosphatase from human erythrocytes. Arch Biochem. Biophys. 313, 166- 172 https://doi.org/10.1006/abbi.1994.1373
  13. Glenn, G. M., Krober, M. S., Kelly, P., McCarty, J. and Weir, M. (1995) Pyridoxine as therapy in theophyline-induced seizure. Ver. Hum. Nerol. 395, 177-194
  14. Gospe Jr, S. M., Olin, K. L. and Keen, C. L. (1994) Reduced GABA synthesis in pyridoxine dependent seizures. Lancet 343, 1133-1134 https://doi.org/10.1016/S0140-6736(94)90236-4
  15. Hanna, M. C., Turner, A. J. and Kirkness, E. F. (1997) Human pyridoxal kinase. J. Biol. Chem. 272, 10756-10760 https://doi.org/10.1074/jbc.272.16.10756
  16. Harris, H. (1990) The human alkaline phosphatases: what we know and what we don't know. Clin. Chim. Acta 186, 133- 150 https://doi.org/10.1016/0009-8981(90)90031-M
  17. Jang, Y. M., Kim, D. W., Kang, T. C., Won, M. H., Baek, N. I., Moon, B. J., Choi, S. Y. and Kwon, O. S. (2003) Human pyridoxal phosphatase: Molecular cloning, functional expression, and tissue distribution. J. Biol. Chem. 278, 50040- 50046 https://doi.org/10.1074/jbc.M309619200
  18. Kwok, F. and Churchich, J. E. (1992) Pyridoxine-5'-P oxidase; in Chemistry and Biochemistry of Flavoenzymes, Muller, F. (ed.) pp. 1-20, CRC Press, London, UK
  19. Kwon, O. S., Kwok, F. and Churchich, J. E. (1991) Catalytic and regulatory properties of native and chymotrypsin-treated pyridoxine-5-phosphate oxidase. J. Biol. Chem. 266, 22136- 22140
  20. Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680- 685 https://doi.org/10.1038/227680a0
  21. Lee, H. Y., Eum, W. S., Kim, D. W., Lee, B. R., Yoon, C. S., Jang, S. H., Choi, H. S., Choi, S. H., Back, N. I., Kang, J. H., Kang, T. C., Won, M. H., Cho, S. W., Lee, K. S., Park, J. and Choi, S. Y. (2003) Isolation and identification of antioxidant enzyme catalase stimulatory compound from Garnoderma lucidum. J. Biochem. Mol. Biol. 36, 450-455 https://doi.org/10.5483/BMBRep.2003.36.5.450
  22. Li, T. K., Lumeng, L. and Veitch, R. L. (1974) Regulation of pyridoxal-5'-phosphate metabolism in liver. Biochem. Biophys. Res. Commun. 61, 677-684 https://doi.org/10.1016/0006-291X(74)91010-9
  23. Lumeng, L. and Li, T. K. (1975) Characterization of the pyridoxal- 5'-phosphate and pyridoxamine-5'-phosphate hydrolase activity in rat liver: Identity with alkaline phosphatase. J. Biol. Chem. 250, 8126-8131
  24. Malmqvist, M. (1993) Biospecific interaction analysis using biosensor technology. Nature 361, 186-187 https://doi.org/10.1038/361186a0
  25. McCormick, D. B., Gregory, M. E. and Snell, E. E. (1961) Pyridoxal phosphokinase I: assay, distribution, purification and properties. J. Biol. Chem. 236, 2076-2084
  26. Meister, A. (1990) On the transamination of enzymes. Ann. N. Y. Acad. Sci. 585, 13-31 https://doi.org/10.1111/j.1749-6632.1990.tb28038.x
  27. Ngo, E. O., LePage, G. R., Thanassi, J. W., Meisler, N. and Netter, L. M. (1998) Absence of pyridoxine-5'-phosphate oxidase (PNPO) activity in neoplastic cells: isolation, characterization, and expression of PNPO cDNA. Biochemistry 37, 7741-7748 https://doi.org/10.1021/bi972983r
  28. Snell, E. E. (1990) Vitamine $B_6$ and decarboxylation of histidine. Ann. N. Y. Acad. Sci. 585, 1-12 https://doi.org/10.1111/j.1749-6632.1990.tb28037.x
  29. Snell, E. E. and Haskell, B. E. (1971) The metabolism of vitamine $B_6$; in Comprehensive Biochemistry, Florkin, M., and Stotz, E. H., (eds.), pp. 47-71, Elsevier Scientific Publishing Co., Netherlands
  30. Towbin, H., Staehelin, T. and Gordon, J. (1978) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedures and some applications. Proc. Natl. Acad. Sci. USA 76, 7350-4354
  31. Waymire, K. G., Mahuren, J. D., Jaje, J. M., Guilarte, T. R., Coburn, S. P. and MacGregor, G. R. (1995) Mice lacking tissues non-specific alkaline phosphatase die from seizures due to defective metabolism of vitamin $B_6$. Nat. Genet. 11, 45-51 https://doi.org/10.1038/ng0995-45

Cited by

  1. Pyridoxal-5′-phosphate phosphatase/chronophin inhibits long-term potentiation induction in the rat dentate gyrus vol.19, pp.11, 2009, https://doi.org/10.1002/hipo.20568
  2. Potential role of pyridoxal-5′-phosphate phosphatase/chronopin in epilepsy vol.211, pp.1, 2008, https://doi.org/10.1016/j.expneurol.2008.01.029