Phosphatidylcholine de novo synthesis and modification are carried out sequentially in HL60 cells: evidence from mass isotopomer distribution analysis.
The traditional (parallel) model of molecular species synthesis of phosphatidylcholine is based on the substrate specificity of two glycerolphosphate acyltransferases. Preformed molecular species of diacylglycerols are then converted to phosphatidylcholine. In this investigation, we used [1,2,3,4-(13)C(4)]palmitate as a tracer to determine the turnover rates of diacylglycerols and phosphatidylcholines. In HL60 cells, the fractional turnover rate is 34.1 +/- 16.6%/h for 1,2-dipalmitoylglycerophosphocholine (16:0,16:0-GPC), which accounts for approximately 10% of total diacylglycerol turnover. The turnover rates of other phosphotidylcholines reflect the primary event of 16:0,16:0-GPC turnover. In addition, the distribution of mass isotopomers is used to study the biosynthesis of diacylglycerols and phosphatidylcholines. On the basis of precursor-product enrichments, we propose a sequential model to account for the synthesis of phosphatidylcholine molecular species. In this model, 1,2-dipalmitoylglycerol is the only molecular species used for the synthesis of phosphatidylcholine. This precursor is converted to 1,2-dipalmitoylglycerophosphocholine, which is then deacylated to provide substrates for chain elongation and/or desaturation. These modified acyl substrates are then reacylated back to form other molecular species. This sequential model is consistent with palmitate being the dominant fatty acid product derived from mammalian fatty acid synthase. It has the advantage of protecting cells from acyl modification by exogenous substrates. Furthermore, this sequence generates only inert 1,2-dipalmitoylglycerol instead of the active diacylglycerol molecular species that contain unsaturated fatty acids.
Tserng
Kou-Yi
KY
Medical Research Service, Veterans Affairs Medical Center, and Department of Nutrition, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106, USA. kxt3@cwru.edu
Griffin
Ronda L
RL
D002247:Carbon Isotopes; D018929:Cell Culture Techniques / Q000379:methods; D003470:Culture Media / Q000737:chemistry; D004075:Diglycerides / Q000737:chemistry / Q000378:metabolism; D005227:Fatty Acids / Q000096:biosynthesis / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D018922:HL-60 Cells; D006801:Humans; D019301:Oleic Acid / Q000378:metabolism; D010168:Palmitates / Q000737:chemistry / Q000378:metabolism; D010713:Phosphatidylcholines / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism
D002247:Carbon Isotopes; D003470:Culture Media; D004075:Diglycerides; D005227:Fatty Acids; D010168:Palmitates; D010713:Phosphatidylcholines; D019301:Oleic Acid