title: Reconstitution of phosphatidylserine transport from chemically defined donor membranes to phosphatidylserine decarboxylase 2 implicates specific lipid domains in the process.
abstract: Phosphatidylserine (PtdSer) is transported from its site of synthesis in the endoplasmic reticulum to the locus of PtdSer decarboxylase 2 (Psd2p) in the Golgi/vacuole and decarboxylated to form phosphatidylethanolamine. Recent biochemical and genetic evidence has implicated the C2 domain of Psd2p and a membrane-bound form of the phosphatidylinositol binding/transfer protein, PstB2p, as essential for this transport process. We devised a reconstituted system in which chemically defined donor membranes function to transfer PtdSer to the biological acceptor membranes containing Psd2p. The transfer of PtdSer is poor when the donor membranes have a high degree of curvature but markedly enhanced when the membranes are relatively planar (> or =400-nm diameter). PtdSer transfer is also dependent upon both the bulk and the surface concentrations of the lipid, with pure PtdSer vesicles acting as the most efficient donors at all concentrations. The lipid transfer from donor membranes containing either 100% PtdSer or 50% PtdSer at a fixed concentration (e.g. 250 microM PtdSer) differs by a factor of 20. Surface dilution of PtdSer by choline, ethanolamine, glycerol, and inositol phospholipids markedly inhibits PtdSer transfer, whereas phosphatidic acid (PtdOH) stimulates the transfer. Most importantly, the transfer of PtdSer from liposomes to Psd2p fails to occur in acceptor membranes from strains lacking PstB2p or the C2 domain of Psd2p. These data support a model for PtdSer transport from planar domains highly enriched in PtdSer or in PtdSer plus PtdOH.
authors.affiliation: Program in Cell Biology, Department of Medicine, National Jewish Medical and Research Center, Denver, Colorado 80206, USA.
authors.forename: Wen-I
authors.initials: WI
authors.lastname: Wu
authors.forename: Dennis R
authors.initials: DR
authors.lastname: Voelker
mesh_terms: D001692:Biological Transport; D002262:Carboxy-Lyases / Q000737:chemistry*; D002462:Cell Membrane / Q000378:metabolism; D002712:Chlorides / Q000494:pharmacology; D002794:Choline / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D019856:Ethanolamine / Q000737:chemistry; D005656:Fungal Proteins / Q000378:metabolism; D005990:Glycerol / Q000737:chemistry; D007700:Kinetics; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D017895:Manganese Compounds / Q000494:pharmacology; D008567:Membranes, Artificial; D010712:Phosphatidic Acids / Q000737:chemistry; D010716:Phosphatidylinositols / Q000737:chemistry; D010718:Phosphatidylserines / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D015003:Yeasts / Q000502:physiology
chemical_list: D002712:Chlorides; D005656:Fungal Proteins; D008055:Lipids; D008081:Liposomes; D017895:Manganese Compounds; D008567:Membranes, Artificial; D010712:Phosphatidic Acids; D010716:Phosphatidylinositols; D010718:Phosphatidylserines; D019856:Ethanolamine; D002262:Carboxy-Lyases; C022353:phosphatidylserine decarboxylase; D002794:Choline; D005990:Glycerol; C025340:manganese chloride