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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Purification and Characterization of Protein Methylase II from Porcine Testis

  • Jung, Ki-Kyung (Department of Genetic Engineering, College of Life Science and Natural Resources, Sungkyunkwan University) ;
  • Kwon, Myung-Hee (Department of Genetic Engineering, College of Life Science and Natural Resources, Sungkyunkwan University) ;
  • Lee, Hoi-Young (Laboratory of Pathology, National Cancer Institute, National Institute of Health) ;
  • Lee, Hyang-Woo (Laboratory of Biochemistry, College of Pharmacy, Sungkyunkwan University) ;
  • Hong, Sung-Youl (Department of Genetic Engineering, College of Life Science and Natural Resources, Sungkyunkwan University)
  • Received : 1994.10.15
  • Published : 1995.03.31
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Abstract

Protein methylase II (S-adenosyl-L-methionine : protein O-methyl-transferase, EC 2.1.1.24; PM II) was purified approximately 1250-fold from porcine testis by fractional precipitation and DEAE-cellulose chromatography, followed by gel filtration on a Sephadex G-75 column and HPLC on a Protein Pak 125 column. The molecular weight of the enzyme was estimated to be 33,000 daltons by SDS-PAGE, which agreed with the value determined by gel filtration. Isoelectric focusing of purified PM II showed a single protein species with an isoelectric point of 6.2. The optimum pH for the reaction was 6.0. The $K_m$ value of the enzyme was $1{\times}10^{-5}M$ with a $V_{max}$ value of 769 pmol/min/mg of enzyme. S-adenosyl-L-homocysteine is a competitive inhibitor of PM II with a $K_i$ value of $1.38{\times}10^{-6}M$.

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