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The Pharmaceutical Society of Korea (대한약학회)
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Polymerization of ADP-Ribose Pyrophosphatase: Conversion Mechanism of $Mg^{2+}-Dependent$ ADP-Ribose Pyrophosphatase into $Mg^{2+}-Independent$ Form

  • Kim, Dae-Ki (Microbiology and Immunology, Chonbuk National University Medical School) ;
  • Kim, Jong-Hyun (Obstetrics and Gynecology, Chonbuk National University Medical School) ;
  • Song, Eun-Kyung (Department of Biochemistry, Institute of Medical Science, Microbiology and Immunology, Chonbuk National University Medical School) ;
  • Han, Myung-Kwan (Microbiology and Immunology, Chonbuk National University Medical School) ;
  • Kim, Jong-Suk (Department of Biochemistry, Institute of Medical Science, Chonbuk National University Medical School)
  • Published : 2003.10.01
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Abstract

ADP-ribose pyrophosphatase (ADPRase) hydrolyzes ADP-ribose (ADPR) into AMP and ribose-5'-phosphate. It is classified into two groups, $Mg^{2+}$-dependent and $Mg^{2+}$-independent ADPRase, depending on its $Mg^{2+}$requirement. Here, we purified $Mg^{2+}$-dependent ADPRase from rabbit liver and examined what factors affect $Mg^{2+}$ requirement. The purified enzyme showed a single band with the molecular weight of 34 kDa on SDS-PAGE both in the presence and absence of 2-mercaptoethanol. The molecular weight of the native enzyme calculated by gel filtration was 68 kDa, indicating that ADPRase is a dimer made up of two identical subunits. $Mg^{2+}$-dependent ADPRase with the highest ADPR affinity had a $K_m$ of 160$\pm$10 $\mu$M and a pH optimum of around pH 9.5. Treatment of the purified ADPRase with heated cytosol fractions at 37$^{\circ}C$ for 3 h caused some changes in the chemical properties of the enzyme, including an increase in molecular weight, a decrease in solubility, and a loss of $Mg^{2+}$-dependency. The molecular weight of the cytosol-treated ADPRase measured by gel filtration was over 420 kDa, suggesting, for the first time, that ADPRase could be polymerized by undefined cytoplasmic factors, and that polymerization is accompanied by changes in the solubility and metal ion dependency of the enzyme.

Keywords

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