Journal of the Korean Magnetic Resonance Society (한국자기공명학회논문지)

Korean Magnetic Resonance Society (한국자기공명학회)
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Purification and NMR studies on Phosphatase domain of UBLCP1

  • Oh, Hyo-Sun (Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University) ;
  • Ko, Sung-Geon (Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University) ;
  • Moon, Sun-Jin (Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University) ;
  • Shin, Hang-Cheol (Department of Bioinformatics and Life Science, Soongsil University) ;
  • Lee, Weon-Tae (Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University)
  • Published : 2009.12.20
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Abstract

UBLCP1 is composed of Ubiquitin Like domain and RNA Polymerase II Phosphatase I domain. Phosphatase domain (25.9KDa) has been cloned into the E.coli using pET32a vector with TEV protease cleavage site and successfully purified as a monomer using affinity chromatography and histidine tag was cleaved with TEV protease for structural studies. Our results indicated that the Phosphatase domain showed well-defined folded structure based on data from one-dimensional and two-dimensional NMR spectroscopy. Data form circular dichroism also suggested that Phosphatase domain consisted of both ${\alpha}$ -helix and ${\beta}$ -sheet. This information will be used for detailed structural study of UBLCP1.

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References

  1. Ko S.G., Lee Y.M., Yoon J.B., Lee W.T., Bulletin of the Korean Chemical Society 30, No.5 (1039)
  2. Zheng H., Ji C., Gu S., Shi B., Wang J., Xie Y., Mao Y., Biochem Biophys Res Commun. 331(4), 1401-7 (2005) https://doi.org/10.1016/j.bbrc.2005.04.065
  3. Michael S., Archambault J., William L., Frank C. P. H., Gileadi O., David B., Ezra G. Jennings, Kouyoumdjian F., Alan R.D., Richard A.Y., Greenblatt J., molecular cell. 4, Issue 1 Pages 55-62 (1999) https://doi.org/10.1016/S1097-2765(00)80187-2
  4. Michele Y., Patrick S., Dahmus Michael E., Gordon N., J. Biological Chem. 278, 26078-26085 (2003) https://doi.org/10.1074/jbc.M301791200
  5. ZAWEL L., REINBERG D., Progress in nucleic acid research and molecular biology 44, 67-108.Stephen B.(1993) https://doi.org/10.1016/S0079-6603(08)60217-2
  6. THOMPSON C. M., KOLESKE A. J., CHAO D. M., YOUNG R. A., J. Cell Sci. Cell ISSN 0092-8674, 73, no7, 1361-1375 (1993) https://doi.org/10.1016/0092-8674(93)90362-T
  7. Stephen B., Nature structural biology 10, number9 (2003)
  8. Yunokuchi, , Fan H., Iwamoto Y., Araki C., Yuda M., Umemura H., Harada F., Ohkuma Y., Hirose Y., GENES CELLS. 14, 1105-1118 (2009) https://doi.org/10.1111/j.1365-2443.2009.01339.x
  9. Provencher, Juergen G., Biochemistry 20(1), 33- 37 (1981) https://doi.org/10.1021/bi00504a006
  10. Thole B. T., Carra P., Sette F., van der Laan G., Phys. Rev. Lett. 68, 1943 - 1946 (1992) https://doi.org/10.1103/PhysRevLett.68.1943
  11. Radek M., Luka K., Vladimir S., Tetrahedron Letters 38, Issue 4 665-668 (1997) https://doi.org/10.1016/S0040-4039(96)02387-8