KSBB Journal

  • 제21권1호
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  • Pages.16-19
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  • 2006
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  • 1225-7117(pISSN)
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  • 2288-8268(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

2-D 전기영동 분석을 통한 $H_2O_2$와 연계된 효모 시스템 NDPK에 관한 연구

Two-dimensional Electrophoretic Analysis of Nucleotide phosphate Kinase Mediated Hydrogen Peroxide Cross-linking in Saccharamyces cerevisiae

  • 문혜정 (경희대학교 산학협력기술연구원) ;
  • 윤대진 (경상대학교 대학원 응용생명과학부) ;
  • 박창호 (경희대학교 환경.응용화학대학)
  • Moon Hae-Jeong (Industrial Liaison Research Institute) ;
  • Yun Dae-Jin (Department of Chemical Engineering, Kyung Hee University) ;
  • Park Chang-Ho (Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University)
  • 발행 : 2006.02.01
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초록

최근의 연구에 의하면 열이나, 산화적 스트레스에 대해서 NDPK는 구조적인 변화를 유발하며, 효소 활성과 구조가 oxidant에 의해 변화된다는 보고를 근거로 하여 정상적인 효모균주와 효모의 NDPK 유전자가 파괴된 mutant에서, 산화적 스트레스에 관련된 역할을 규명하고자 2-D 전기영동 방법을 통해서, $H_2O_2$의 처리전과 처리 후에 전사패턴이 변화된 유전자들, 즉, 산화적조절 신호체제에 연관되어졌을 것이라고 생각되어지는 몇 개의 단백질 리스트를 얻었다. 이 결과는 NDPK의 redox state의 조절에 관련된 효소의 성질을 규명함에 있어 유용한 유전자 신호 체제정보를 제공할 것으로 생각되어진다.

Oxidative modification of nucleoside diphosphate kinase (NDPK) is identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometer. The quaternary structure of NDPK appears to be regulated by cross-linking with an oxidant, $H_2O_2$. We compared roles of NDPK in each of wild type and ynk mutant against oxidative stress. Six specific proteins changed by $H_2O_2$ were identified using two-dimensional electrophoretic analysis. YNK regulated several proteins, related to $H_2O_2$ signaling functions. These results suggest that one of the important functions of NDPK is the regulation of cellular redox state.

키워드

참고문헌

  1. Kumagai, Y., T. Wakayama, S. Lib, A. Shinohara, A. Iwamatsu, G. Sun, and N. Shimojo (2000), Zeta-crystallin catalyzes the reductive activation of 2,4,6-trinitrotoluene to generate reactive oxygen species: a proposed mechanism for the induction of cataracts, FEES Lett. 478, 295-298 https://doi.org/10.1016/S0014-5793(00)01841-X
  2. Tabatabaie, T. and T. A. Floyd (1996), Inactivation of glutathione peroxidase by benzaldehyde, Toxicol. Appl. Pharmacal. 141, 389-393 https://doi.org/10.1006/taap.1996.0304
  3. Shen, Y., H. M. Shen, C. Y. Shi, and C. N. Ong (1996), Benzene metabolites enhance reactive oxygen species generation in HL60 human leukemia cells, Hum. Exp. Toxicol. 15, 422-427 https://doi.org/10.1177/096032719601500510
  4. Mesnildrey, S., F. Agou, and M. Vero (1997), The in vitro DNA binding properties of NDP kinases are related to its oligomeric state, FEBS Lett. 418, 53-57 https://doi.org/10.1016/S0014-5793(97)01292-1
  5. Leung, S. M. and L. F Hightower (1997), A 16-kDa protein functions as a new regulatory protein for Hsc70 molecular chaperone and is identified as a member of the nm23/nucleoside diphosphate kinase family, J. Biol. Chem. 272, 2607-2614 https://doi.org/10.1074/jbc.272.5.2607
  6. Song, E. J., Y. S. Kim, J. Y. Chung, E. Kim, S. K. Chae, and K. J. Lee (2000), Oxidative modification of nucleoside diphosphate kinase and its identification by Matrix Assisted Laser Desorption/ Ionization time of flight mass spectrophotometry, Biochemistry 39, 10090-10097 https://doi.org/10.1021/bi000267a
  7. Chakrabarthy, A. M. (1998), Nucleoside diphosphate kinase: role in bacterial growth, virulence, cell signaling and polysaccharide synthesis, Mol. Microbiol. 28, 875-882 https://doi.org/10.1046/j.1365-2958.1998.00846.x
  8. Cippollini, G., A. Berti, L. Fiore, G. Rainaldi, F. Basolo, G. Merlo, G. Beveilaqua, and M. Galigo (1997), Down regulation of the nm23.h1 gene inhibits cell proliferation, Int. J. Cancer 73, 297-302 https://doi.org/10.1002/(SICI)1097-0215(19971009)73:2<297::AID-IJC22>3.0.CO;2-B
  9. Engel, M., M. Seifert, B. Theisinger, U. Seyfert, and C. Welter (1998), Glyceraldehyde-3-phosphate dehydrogenase and NM23-H1/nucleside diphosphate kinase A: two old enzymes for the novel Nm23 protein phosphotransferase function, J. Biol. Chem. 273, 20058-20065 https://doi.org/10.1074/jbc.273.32.20058
  10. Postel, E. H., S. J. Berberich, J. Flint, and C. A. Ferone (1993), Human c-myc transcription factor PuF identified as nm23-H2 nucleoside diphosphate kinase, a candidate suppressor of tumour metastasis, Science 261, 478-480 https://doi.org/10.1126/science.8392752
  11. Choi, G., H. Yi, J. Lee, Y. K. Kwon, M. S. Soh, B. Shin, Z. Luka, T. R. Hahn, and P. S. Song (1999), Phytochrome signaling is mediated through nucleoside diphosphate kinase2, Nature 401, 610-613 https://doi.org/10.1038/44176
  12. Zimmermann, S., A. Baumann, K. Jaekel, I. Marbach, D. Engelberg, and H. Frohnmeyer (1999), UV resposnsive genes of arabidopsis revealed by similarity to the Gcn4-mediated UV response in yeast, J. Biol. Chem. 274, 17017-17024 https://doi.org/10.1074/jbc.274.24.17017
  13. Escobar-Galvis, M. L., S. Martilla, G. Hakansson, J. Forseberg, and C. Knorpp (2001), Heat stress response in pea involves interaction of mitochondrial nucleoside diphosphate kinase with a novel 86 kD protein, Plant Physiol. 126, 69-77 https://doi.org/10.1104/pp.126.1.69
  14. Barthel, T. K. and G. C. Walker (1999), Inferences concerning the ATPase properties of DnaK and other HSP70s are affected by the ADP kinase activity of the co-purifying nucleoside diphosphate kinase, J. Biol. Chem. 274, 36670-36678 https://doi.org/10.1074/jbc.274.51.36670
  15. Davidson, J. F., B. Whyte, P. H. Bissinger, and R. H. Schiestl (1996), Oxidative stress is involved in heat-induced cell death in S. cerevisiae, Proc. Natl. Acad. Sci. USA 93, 5116-5121
  16. Sherman, F. (1991), Getting started with yeast, Met. Enzymol. 194, 12-17
  17. Elble, R. (1992), A simple and an efficient procedure for transformation of yeast, BioTechniques 13, 18-22
  18. Kim, S. T., K. S. Cho, Y. S. Jang, and K. Y. Kang (2001), Two-dimensional electrophoretic analysis of rice proteins by polyethylene glycol fractionation for protein arrays, Electrophoresis 22, 2103-2109 https://doi.org/10.1002/1522-2683(200106)22:10<2103::AID-ELPS2103>3.0.CO;2-W
  19. Vorum, H., M. Ostergaard, P. Hensechke, J. J Enghild, M. Riazati, and G. E. Rice (2004), Proteomic analysis of hyperoxia-induced responses in the human choriocarcinoma cell line JEG-3, Proteomics 4. 861-867 https://doi.org/10.1002/pmic.200300639