Activation of polyisoprenyl diphosphate phosphatase 1 remodels cellular presqualene diphosphate.
Polyisoprenyl diphosphates play diverse and vital roles in cell function in health and disease. The counter-regulatory lipid signaling molecule, presqualene diphosphate (PSDP), is rapidly converted to its monophosphate form (PSMP) upon cell activation [Levy, B. D., Petasis, N. A., and Serhan, C. N. (1997) Nature 389, 985-990]. The first PSDP phosphatase was recently identified and named polyisoprenyl diphosphate phosphatase 1 (PDP1) [Fukunaga, K., et al. (2006) J. Biol. Chem. 281, 9490-9497]. Here, we present evidence that PDP1 displays properties of a lipid phosphate phosphatase/phosphotransferase with distinct substrate preference for PSDP. Cell activation with PMA increased PSDP phosphatase activity in a concentration-dependent manner, and Western analysis suggested that PDP1 is directly phosphorylated by protein kinase C. Cellular PSDP phosphatase activity was also induced by the receptor-mediated agonists insulin and TNF-alpha. To address PDP1's contribution to cellular PSDP phosphatase activity, HEK293 cells were established that stably expressed PDP1 siRNA, leading to a 60% decrease in the level of PDP1 RNA, and concomitant decreases in PDP1 protein and PMA-initiated PSDP phosphatase activity. HEK293 cells harboring the PDP1 siRNA construct also displayed a marked decrease in the extent of PMA-initiated conversion of cellular PSDP to PSMP. Together, these findings are the first to indicate that PDP1 is activated during cell responses to soluble stimuli to convert PSDP to PSMP. Moreover, they provide evidence that PDP1 can serve as a new checkpoint for polyisoprenyl phosphate remodeling during cell activation.
Carlo
Troy
T
Pulmonary and Critical Care Medicine, Brigham and Women's Hospital and Harvard Medical School, 75 Francis Street, Boston, Massachusetts 02115, USA.
Petasis
Nicos A
NA
Levy
Bruce D
BD
D002460:Cell Line; D004789:Enzyme Activation; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D018716:Molecular Mimicry; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D013379:Substrate Specificity
D008565:Membrane Proteins; D011106:Polyisoprenyl Phosphates; D034741:RNA, Small Interfering; C017672:presqualene pyrophosphate; C401298:sphingosine-1-phosphate phosphatase; D010744:Phosphoric Monoester Hydrolases