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The Pharmaceutical Society of Korea (대한약학회)
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Identification of Three Competitive Inhibitors for Membrane­Associated, $Mg^{2+}-Dependent$ and Neutral 60 kDa Sphingomyelinase Activity

  • Kim Seok Kyun (Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Jung Sang Mi (Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Ahn Kyong Hoon (Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Jeon Hyung Jun (Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Lee Dong Hun (Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Jung Kwang Mook (Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Jung Sung Yun (Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Kim Dae Kyong (Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University)
  • Published : 2005.08.01
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Abstract

Methanol extracts of domestic plants of Korea were evaluated as a potential inhibitor of neutral pH optimum and membrane-associated 60 kDa sphingomyelinase (N-SMase) activity. In this study, we partially purified N-SMase from bovine brain membranes using ammonium sulfate. It was purified approximately 163-fold by the sequential use of DE52, Butyl-Toyopearl, DEAE-Cellulose, and Phenyl-5PW column chromatographies. The purified N-SMase activity was assayed in the presence of the plant extracts of three hundreds species. Based on the in vitro assay, three plant extracts significantly inhibited the N-SMase activity in a time- and concentration-dependent manner. To further examine the inhibitory pattern, a Dixon plot was constructed for each of the plant extracts. The extracts of Abies nephrolepis, Acer tegmentosum, and Ginkgo biloba revealed a competitive inhibition with the inhibition constant (Ki) of $11.9 {\mu}g/mL,\;9.4{\mu}g/mL,\;and\;12.9{\mu}g/mL$, respectively. These extracts also inhibited in a dose-dependent manner the production of ceramide induced by serum deprivation in human neuroblastoma cell line SH-SY5Y.

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References

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