We reported the presence of group V secretory phos- pholipase A2 (GV) in the nuclei of cultured neuronal cells whereas group IIA (GIIA) was largely found in mitochondria of the same cells and in rat brain cortex mitochondria (Macchioni et al., 2004). In this study, PLA2 activity was assayed in intact and lysed rat brain cortex nuclei using [3H]oleate labelled E. coli mem- branes as substrate and we observed that the specific activity was higher in nuclear lysate than in intact nuclei. DTT or Indoxam (Suzuki et al., 2000), inhibitors of sPLA2s, greatly reduced enzyme activity. The addition of AACOCF3 or MAFP, inhibitors of cPLA2s were inef- fective. BEL, inhibitor of GVI PLA2 (Lucas et al., 2005) slightly reduced PLA2 activity in intact nuclei. PLA2 activity was also monitored by fluorescence microscopy using the fluorogenic ethanolamine glycerophospholipid (PED6) as substrate (Shinzawa et al., 2003). These exper- iments confirmed the presence of a DTT-sensitive PLA2 in nuclear membrane that was identified by Western blot analysis as GV in glial and neuronal nuclei puri- fied from rat brain cortex. In the same fractions GIIA was undetectable. We conclude that PLA2 activity of nuclei of neural cells has to be mainly attributed to GV. Acknowledgment: Supported by grant MIUR-PRIN 2004. References: Macchioni, L., et al., 2004. J. Biol. Chem. 279, 37860. Suzuki, N., et al., 2000. J. Biol. Chem. 275, 5785. Lucas, K.K., et al., 2005. Prostagland. Lipid Mediators 77, 235. Shinzawa, K., et al., 2003. J. Cell. Biol. 163, 1219.
Presence of Group V Phospholipase A2 in rat brain cortex nuclei
NARDICCHI, Vincenza;MACCHIONI, Lara;FERRINI, Monica;BIANCONI, Marcello;GORACCI, Gianfrancesco
2006
Abstract
We reported the presence of group V secretory phos- pholipase A2 (GV) in the nuclei of cultured neuronal cells whereas group IIA (GIIA) was largely found in mitochondria of the same cells and in rat brain cortex mitochondria (Macchioni et al., 2004). In this study, PLA2 activity was assayed in intact and lysed rat brain cortex nuclei using [3H]oleate labelled E. coli mem- branes as substrate and we observed that the specific activity was higher in nuclear lysate than in intact nuclei. DTT or Indoxam (Suzuki et al., 2000), inhibitors of sPLA2s, greatly reduced enzyme activity. The addition of AACOCF3 or MAFP, inhibitors of cPLA2s were inef- fective. BEL, inhibitor of GVI PLA2 (Lucas et al., 2005) slightly reduced PLA2 activity in intact nuclei. PLA2 activity was also monitored by fluorescence microscopy using the fluorogenic ethanolamine glycerophospholipid (PED6) as substrate (Shinzawa et al., 2003). These exper- iments confirmed the presence of a DTT-sensitive PLA2 in nuclear membrane that was identified by Western blot analysis as GV in glial and neuronal nuclei puri- fied from rat brain cortex. In the same fractions GIIA was undetectable. We conclude that PLA2 activity of nuclei of neural cells has to be mainly attributed to GV. Acknowledgment: Supported by grant MIUR-PRIN 2004. References: Macchioni, L., et al., 2004. J. Biol. Chem. 279, 37860. Suzuki, N., et al., 2000. J. Biol. Chem. 275, 5785. Lucas, K.K., et al., 2005. Prostagland. Lipid Mediators 77, 235. Shinzawa, K., et al., 2003. J. Cell. Biol. 163, 1219.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
