BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Identification of Phospholipase C Activated by $GTP{\gamma}S$ in Plasma Membrane of Oat Cell

  • Kim, Hyae-Kyeong (Department of Biochemistry, Chungbuk National University) ;
  • Park, Moon-Hwan (Department of Biochemistry, Chungbuk National University) ;
  • Chae, Quae (Department of Biochemistry, Chungbuk National University)
  • Received : 1995.02.20
  • Published : 1995.09.30
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Abstract

In order to investigate whether phospholipase C (PLC) activity in oat celIs is regulated by Gprotein, we have characterized PLC in plasma membranes of oat tissues. To identify the purified plasma membrane, $K^+$-stimulated, $Mg^{2+}$-dependent ATPase activity was measured. The activity of ATPase was shown to be proportional to the concentration of membrane protein. To examine the PLC activity regulated by G-protein, we used the inside-out and outside-out plasma membrane mixture isolated from the oat cells. The plasma membrane mixture showed higher PLC activity than the one of the outside-out plasma membrane. This suggests that PLC activity is located at the cytoplasmic surface of plasma membrane. PLC activity in plasma membrane mixture was dependent on $Ca^{2+}$ with maximum activity at 100 ${\mu}m$ $Ca^{2+}$ and it was inhibited by 1 mM EGTA. Using Sep-pak $Accell^{TM}$ Plus QMA chromatography, we found that inositol 1,4,5-trisphosphate ($IP_3$) was produced in the presence of 10 ${\mu}m$ $Ca^{2+}$. The PLC activity in the membrane was enhanced by an activator of G-protein ($GTP{\gamma}S$) and not by an inhibitor ($GDP{\beta}S$). This indicates that a G-protein is involved in the activation of PLC in the plasma membrane of oat cells.

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