大鼠肾脏中两种14 kDa磷脂酶A2亚型的检测与纯化：它们在类花生酸合成中的作用。

Detection and purification of two 14 kDa phospholipase A2 isoforms in rat kidney: their role in eicosanoid synthesis.

作者信息

Hara S, Kudo I, Komatani T, Takahashi K, Nakatani Y, Natori Y, Ohshima M, Inoue K

机构信息

Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.

出版信息

Biochim Biophys Acta. 1995 Jun 27;1257(1):11-7. doi: 10.1016/0005-2760(95)00011-z.

DOI:10.1016/0005-2760(95)00011-z

PMID:7599175

Abstract

Phospholipase A2 (PLA2) activity in the soluble fraction of rat kidney yielded three peaks on DEAE cellulose column chromatography. From these three, we purified two PLA2 isoforms to near-homogeneity. Both had a molecular weight of approx. 14,000 on SDS-PAGE, and immunochemical and enzymological studies indicated that one is a 14 kDa type I PLA2 and the other a 14 kDa type II PLA2. RNA blot analysis confirmed that rat kidney contains both types of PLA2 and that administration of lipopolysaccharides and mercury chloride into rats increased type II PLA2 mRNA levels in kidney. When cultured rat mesangial cells were incubated with purified type I or type II PLA2 in combination with the calcium ionophore A23187 at suboptimal condition, augmentation of prostaglandin E2 production was observed. Type I and type II forms of PLA2 may play a role in arachidonate metabolism in rat kidney.

摘要

大鼠肾脏可溶性部分中的磷脂酶A2（PLA2）活性在DEAE纤维素柱层析上产生了三个峰。从这三个峰中，我们将两种PLA2同工型纯化至接近均一。两者在SDS-PAGE上的分子量约为14,000，免疫化学和酶学研究表明，一种是14 kDa的I型PLA2，另一种是14 kDa的II型PLA2。RNA印迹分析证实，大鼠肾脏含有这两种类型的PLA2，并且向大鼠体内注射脂多糖和氯化汞会增加肾脏中II型PLA2 mRNA的水平。当在次优条件下将培养的大鼠系膜细胞与纯化的I型或II型PLA2与钙离子载体A23187一起孵育时，观察到前列腺素E2产生增加。I型和II型PLA2可能在大鼠肾脏的花生四烯酸代谢中起作用。