title: Biosynthesis of phosphatidylcholine from a phosphocholine precursor pool derived from the late endosomal/lysosomal degradation of sphingomyelin.
abstract: Previous studies suggest that the steps of the CDP- choline pathway of phosphatidylcholine synthesis are tightly linked in a so-called metabolon. Evidence has been presented that only choline that enters cells through the choline transporter, and not phosphocholine administered to cells by membrane permeabilization, is incorporated into phosphatidylcholine. Here, we show that [(14)C]phosphocholine derived from the lysosomal degradation of [(14)C]choline-labeled sphingomyelin is incorporated as such into phosphatidylcholine in human and mouse fibroblasts. Low density lipoprotein receptor-mediated endocytosis was used to specifically direct [(14)C]sphingomyelin to the lysosomal degradation pathway. Free labeled choline was not found either intracellularly or in the medium, not even when the cells were energy-depleted. Deficiency of lysosomal acid phosphatases in mouse or alkaline phosphatase in human fibroblasts did not affect the incorporation of lysosomal [(14)C]sphingomyelin-derived [(14)C]phosphocholine into phosphatidylcholine, supporting our finding that phosphocholine is not degraded to choline prior to its incorporation into phosphatidylcholine. Inhibition studies and analysis of molecular species showed that exogenous [(3)H]choline and sphingomyelin-derived [(14)C]phosphocholine are incorporated into phosphatidylcholine via a common pathway of synthesis. Our findings provide evidence that, in fibroblasts, phosphocholine derived from sphingomyelin is transported out of the lysosome and subsequently incorporated into phosphatidylcholine without prior hydrolysis of phosphocholine to choline. The findings do not support the existence of a phosphatidylcholine synthesis metabolon in fibroblasts.
authors.affiliation: Departments of Pediatrics and Clinical Genetics, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
authors.forename: S M
authors.initials: SM
authors.lastname: Jansen
authors.forename: J E
authors.initials: JE
authors.lastname: Groener
authors.forename: W
authors.initials: W
authors.lastname: Bax
authors.forename: A
authors.initials: A
authors.lastname: Suter
authors.forename: P
authors.initials: P
authors.lastname: Saftig
authors.forename: P
authors.initials: P
authors.lastname: Somerharju
authors.forename: B J
authors.initials: BJ
authors.lastname: Poorthuis
mesh_terms: D000469:Alkaline Phosphatase / Q000172:deficiency; D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004072:Digitonin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D011992:Endosomes / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D006937:Hypercholesterolemia / Q000378:metabolism; D007014:Hypophosphatasia / Q000378:metabolism; D007202:Indicators and Reagents / Q000494:pharmacology; D008077:Lipoproteins, LDL / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism*; D051379:Mice; D009542:Niemann-Pick Diseases / Q000378:metabolism; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D010767:Phosphorylcholine / Q000378:metabolism*; D013109:Sphingomyelins / Q000378:metabolism*; D013997:Time Factors
chemical_list: D007202:Indicators and Reagents; D008077:Lipoproteins, LDL; D010713:Phosphatidylcholines; D013109:Sphingomyelins; D010767:Phosphorylcholine; D000469:Alkaline Phosphatase; D004072:Digitonin