Studies on the separation of phospholipase A_2 preferred the arachidonoyl species of phospholipids

磷脂酶A_2优选花生四烯酰磷脂的分离研究

• 批准号: 62580135
• 负责人: NAKAGAWA Yasuhito
• 金额: $ 0.96万
• 依托单位: Faculty of Pharmaceutical Sciences, Teikyo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62580135/
• 关键词: Macrophage; Arachidonic Acid; Phospholipase; Calcium; カルモデュリン; リン脂質分子種

The effects of calmodulin antagonists on the amount of free fatty acid produced by alveolar macrophages exposed to opsonized zymosan were determined. Opsonized zymosan-induced free arachidonic acid (20:4) production was dramatically suppressed in the presence of W-7 and trifluoperazine without an effect on the production of other fatty acids. Calmodulin antagonists appeared to inhibit phospholipase A_2, since W-7 abolished the liberation of 20:4 from choline glycerophospholipids (CGP) and inositol glycerophospholipids (IGP$. After the challenge with opsonized zymosan, the 20:4 liberation caused by opsonized zymosan was significantly reduced in the absence of extracellular Ca^<2+>, indicating that the release of arachidonic acid required the influx of the extracelluler Ca^<2+> into cell. On the other hand, oleic acid and linoleic acid liberation from alveolar macrophages exposed to opsonized zymosan was not inhibited after the depletion of extracellular Ca^<2+>.Most of phospholipase A_2 activity was present in the cytosol fraction of alveolar macrophages, representing 70% of the total activity. Calcium enhanced the hydrolysis of 16:0-20:4 GPC with cytosolic phospholipase A_2, although the phdrolysis of 16:0-18:1 and 16:0-18:2 species of CGP was not changed by calcium.The purification of cytosolic phospholipase A_2 has been performed by using the hydrophordic column, ion exchange column and gel filtration column. However, phospholipase A_2 which selectively hydrolyzed the arechidonoyl species of phospholipids failed to be isolated.The present studies indicate that phospholipase A_2 in rabbit alveolar macrophages is controlled by Ca^<2+> and calmodulin, and preferentially hydrolyzes arachidonic acid from phospholipids.

测定钙调素拮抗剂对暴露于调理酵母多糖的肺泡巨噬细胞产生的游离脂肪酸的量的影响。调理酵母多糖诱导的游离花生四烯酸（20：4）的生产显着抑制W-7和三氟拉嗪的存在下，对其他脂肪酸的生产没有影响。钙调素拮抗剂对磷脂酶A_2有抑制作用，因为W-7能抑制胆碱甘油磷脂（CGP）和肌醇甘油磷脂（IGP）释放20：4。在用调理酵母多糖刺激后，在细胞外Ca^2+缺乏的情况下，由调理酵母多糖引起的20：4释放显著减少，表明花生四烯酸的释放需要细胞外Ca^2+流入细胞。另一方面，经酵母多糖调理的肺泡巨噬细胞释放油酸和亚油酸的能力在细胞外Ca^2+耗竭后不受抑制，磷脂酶A_2活性主要存在于胞浆中，占总活性的70%。钙对16：0-18：1和16：0-18：2两种胞浆型磷脂酶A_2的水解无影响，但对16：0-20：4型胞浆型磷脂酶A_2的水解有促进作用。本研究表明，兔肺泡巨噬细胞中的磷脂酶A_2受Ca^（2+）和钙调素的控制，优先水解磷脂中的花生四烯酸。

项目成果

Nakagawa,Y.: J.Choromatogr.in press.

Nakakawa,Y.：J.Choromatogr.出版社。

B. R stow; Y. Nakagawa; H. Rabe; K. Waku; D. Kunze: "Species pattern of phosphatidylinositol from lung surfactant and a comparison of the species pattern of phosphatidylinositol and phosphatidylglycerol synthesized de novo in lung microsomal fractions." B

B.R斯托；

Nakagawa,Y.: Arch.Biochem.Biophys.268. inpress (1989)

中川，Y.：Arch.Biochem.Biophys.268。

Y. Nakagawa; K. Waku: "Separation of the molecular species of diacyl and alkenylacyl subclasses of methyl ester of dinitrophenyl ethanolamine glycerophospholipid by high-performance liquid chromatography." J. Chromatogr.(1989)

Y.中川；

Nakagawa,Y.: Biochem.Biophys.Res.Commun.156. 947-953 (1988)

中川，Y.：Biochem.Biophys.Res.Commun.156。