Journal of the Korean Chemical Society (대한화학회지)

Korean Chemical Society (대한화학회)
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Solubilization of Brain Phospholipase D by Taurodeoxycholate: Activational Effect of Some Matal Ions

Taurodeoxycholate에 의한 뇌 포스포리파제 D의 용해: 몇 금속이온의 활성화 효과

  • Choi, Seok Woo (Department of Chemistry and Center for Molecular Catalysis, Seoul National University) ;
  • Choi, Myung Un (Department of Chemistry and Center for Molecular Catalysis, Seoul National University)
  • 최석우 (서울대학교 자연과학대학 화학과,분자촉매 연구센터) ;
  • 최명언 (서울대학교 자연과학대학 화학과,분자촉매 연구센터)
  • Published : 19971200
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Abstract

Microsomal phospholipase D (PLD) in rat brain was solubilized employing 0.2 % taurodeoxycholate in high ionic strength. Phopholipase D activity was determined by measuring product phophatidic acid (PA) using isotope-labelled dipalmitoylphophatidylcholine as a substrate. The solubilized PLD showed an optimal pH of 6.5 and the highest activity at 30$^{\circ}C.$ These properties were similar to those of microsomal PLD before solubilization. The stimulatory effect of oleic acid was observed at the concentration of 4 mM. When effects of metal ions on PLD activity were examined, alkaline earth metals such as $Mg^{2+},\; Ca^{2+},\; Sr^{2+}, \;Ba^{2+}$ promoted the PA production but $Cu^{2+},\; Cd^{2+},\; Al^{3+},\; Ni^{2+},\; V^{5+}$ showed inhibitory effects.

쥐의 미크로좀 포스포리파제 D(PLD)를 센 이온세기 상태에서 0.2% taurodeoxycholate를 사용하여 용해시켰다. 포스포리파제 D의 활성은 기질로 방사성 동위원소로 표지 된 dipalmitoylphophatidylcholine을 사용하여 생성된 phosphatidic acid(PA)를 측정하여 결정하였다. 용해된 PLD의 초적 pH와 온도는 각각 6.5와 30$^{\circ}C$로서 용해되기 전 미크로좀 상태의 PLD와 비슷하였다. 올레산의 활성화 효과는 4 mM 농도에서 관찰되었다. PLD 활성도에 미치는 금속이온 영향을 조사한 결과 $Mg^{2+},\; Ca^{2+},\; Sr^{2+}, \;Ba^{2+}$와 같은 알칼리 토금속은 모두 PA 생성을 촉진시킨 반면, $Cu^{2+},\; Cd^{2+},\; Al^{3+},\; Ni^{2+},\; V^{5+}$는 억제하였다.

Keywords

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