Detection and purification of two 14 kDa phospholipase A2 isoforms in rat kidney: their role in eicosanoid synthesis.
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.
Hara
S
S
Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Kudo
I
I
Komatani
T
T
Takahashi
K
K
Nakatani
Y
Y
Natori
Y
Y
Ohshima
M
M
Inoue
K
K
D000818:Animals; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D015777:Eicosanoids / Q000096:biosynthesis*; D007527:Isoenzymes / Q000302:isolation & purification* / Q000502:physiology; D007668:Kidney / Q000201:enzymology*; D008969:Molecular Sequence Data; D010741:Phospholipases A / Q000302:isolation & purification* / Q000502:physiology; D054467:Phospholipases A2; D051381:Rats; D017207:Rats, Sprague-Dawley
D017931:DNA Primers; D015777:Eicosanoids; D007527:Isoenzymes; D010741:Phospholipases A; D054467:Phospholipases A2