Nitrogen dioxide-induced phosphatidylserine biosynthesis and subcellular translocation in cultured pulmonary artery endothelial cells.
Exposure to nitrogen dioxide (NO2) increases phosphatidylserine (PS) content in the plasma membranes of pulmonary artery endothelial cells (PAEC). We examined whether the increased PS content is associated with increased uptake of L-serine and/or biosynthesis of PS. Exposure to 5 ppm NO2 increased uptake and incorporation of exogenous L-[14C]serine into whole cells, total cellular lipids, phospholipids, and phospholipid subclasses compared to control. Incorporation of L-[14C]serine into the total lipid extracts from isolated plasma membranes, mitochondria, and microsomes from NO2-exposed cells was increased by 45, 32, and 31%, respectively (p < 0.05 for all membranes). Increased incorporation of L-[14C]serine into the total phospholipids of plasma membranes, mitochondria, and microsomes of NO2-exposed cells was increased by 31, 48, and 33%, respectively (p < 0.05 for all membranes). Incorporation of L-[14C]serine into the PS of plasma membranes and microsomes from NO2-exposed cells was increased by 63 and 89%, respectively (p < 0.05 for both membranes). The incorporation of radioactivity from L-[14C]serine into the phosphatidylethanolamine and phosphatidylcholine contents of plasma membranes, mitochondria, and microsomes from NO2-exposed cells was also observed. Exposure of PAEC to NO2 resulted in a significant (p < 0.01) increase in the activity of PS synthase, the serine base-exchange enzyme located in the microsomes of these cells. When L-[14C]serine-prelabeled microsomes were incubated with unlabeled mitochondria from control and NO2-exposed cells, transfer of PS-derived radioactivity from microsomes to mitochondrial phospholipids was observed. These results demonstrate that exposure to NO2 increases uptake and incorporation of exogenous serine as well as intracellular biosynthesis of PS, resulting in increases in the PS content of PAEC and their plasma membranes.
Division of Pulmonary Medicine, University of Florida College of Medicine, Gainesville.
Y D
YD
Li
J M
JM
Patel
E R
ER
Block
D000704:Analysis of Variance; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D046508:Culture Techniques; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007553:Isotope Labeling; D008055:Lipids / Q000032:analysis; D008861:Microsomes / Q000378:metabolism; D009585:Nitrogen Dioxide / Q000633:toxicity*; D010713:Phosphatidylcholines / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism; D010718:Phosphatidylserines / Q000096:biosynthesis* / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D011651:Pulmonary Artery / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D012694:Serine / Q000378:metabolism*; D013552:Swine
D008055:Lipids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D010743:Phospholipids; D012694:Serine; D009585:Nitrogen Dioxide