神经细胞系PC12中膜相关磷脂酶D的活性

Membrane-associated phospholipase D activity in neural cell line PC12.

作者信息

Banno Y, Ito Y, Ojio K, Kanoh H, Nakashima S, Nozawa Y

机构信息

Department of Biochemistry, Gifu University School of Medicine, Japan.

出版信息

J Lipid Mediat Cell Signal. 1996 Sep;14(1-3):237-43. doi: 10.1016/0929-7855(96)00531-7.

DOI:10.1016/0929-7855(96)00531-7

PMID:8906568

Abstract

Stimulation of PC12 cells with carbachol (Cch) or phorbol 12-myristate 13-acetate (PMA) induced [3H]phosphatidylbutanol (PBut) production. Depletion of extracellular Ca2+ abolished the Cch-mediated phospholipase D (PLD) activation, indicating the requirement of Ca2+ influx. Two different types of PLD activity, oleate-dependent and GTP gamma S-dependent, were examined by using exogenous [3H]phosphatidylcholine substrate. PLD activity of the membrane fraction of PC12 cells was highly dependent on oleate and independent of GTP gamma S. This profile is in sharp contrast to that observed in HL60 cells showing the profound GTP gamma S-induced activation of PLD. The oleate-dependent PLD activity of PC12 membrane was inhibited by high concentrations of Ca2+ and Mg2+. These results indicate that Ca2+ may not directly activate PLD but through some Ca(2+)-dependent mechanism(s) in Cch-stimulated cells.

摘要

用卡巴胆碱（Cch）或佛波酯12 - 肉豆蔻酸酯13 - 乙酸酯（PMA）刺激PC12细胞可诱导[3H]磷脂丁醇（PBut）的产生。细胞外Ca2 +的耗尽消除了Cch介导的磷脂酶D（PLD）激活，表明需要Ca2 +内流。通过使用外源性[3H]磷脂酰胆碱底物检测了两种不同类型的PLD活性，即油酸依赖性和GTPγS依赖性。PC12细胞膜部分的PLD活性高度依赖于油酸且不依赖于GTPγS。该特征与在HL60细胞中观察到的情况形成鲜明对比，在HL60细胞中显示出GTPγS诱导的PLD深度激活。PC12细胞膜的油酸依赖性PLD活性受到高浓度Ca2 +和Mg2 +的抑制。这些结果表明，在Cch刺激的细胞中，Ca2 +可能不是直接激活PLD，而是通过一些Ca(2 +)依赖性机制来激活。