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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

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.

Keywords

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