title: Quantitative profiling of the endonuclear glycerophospholipidome of murine embryonic fibroblasts.
abstract: A reliable method for purifying envelope-stripped nuclei from immortalized murine embryonic fibroblasts (iMEFs) was established. Quantitative profiling of the glycerophospholipids (GPLs) in envelope-free iMEF nuclei yields several conclusions. First, we find the endonuclear glycerophospholipidome differs from that of bulk membranes, and phosphatidylcholine (PtdCho) and phosphatidylethanolamine species are the most abundant endonuclear GPLs by mass. By contrast, phosphatidylinositol (PtdIns) represents a minor species. We also find only a slight enrichment of saturated versus unsaturated GPL species in iMEF endonuclear fractions. Moreover, much lower values for GPL mass were measured in the iMEF nuclear matrix than those reported for envelope-stripped IMF-32 nuclei. The collective results indicate that the nuclear matrix in these cells is a GPL-poor environment where GPL occupies only approximately 0.1% of the total nuclear matrix volume. This value suggests GPL accommodation in this compartment can be satisfied by binding to resident proteins. Finally, we find no significant role for the PtdIns/PtdCho-transfer protein, PITPα, in shuttling PtdIns into the iMEF nuclear matrix.
authors.affiliation: Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, NC Emily.k.tribble@gmail.com vytas@tamhsc.edu.
authors.forename: Emily K
authors.initials: EK
authors.lastname: Tribble
authors.affiliation: Departments of Pharmacology and Biochemistry, Vanderbilt University School of Medicine, Vanderbilt Institute of Chemical Biology, Nashville, TN.
authors.forename: Pavlina T
authors.initials: PT
authors.lastname: Ivanova
authors.affiliation: Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, NC Department of Molecular and Cellular Medicine, Texas A&M Health Science Center, College Station, TX.
authors.forename: Aby
authors.initials: A
authors.lastname: Grabon
authors.affiliation: Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, NC.
authors.forename: James G
authors.initials: JG
authors.lastname: Alb
authors.affiliation: Cellular Signaling Laboratory, Department of Biomedical Sciences, University of Bologna, Bologna, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Faenza
authors.affiliation: Cellular Signaling Laboratory, Department of Biomedical Sciences, University of Bologna, Bologna, Italy.
authors.forename: Lucio
authors.initials: L
authors.lastname: Cocco
authors.affiliation: Departments of Pharmacology and Biochemistry, Vanderbilt University School of Medicine, Vanderbilt Institute of Chemical Biology, Nashville, TN.
authors.forename: H Alex
authors.initials: HA
authors.lastname: Brown
authors.affiliation: Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, NC Department of Molecular and Cellular Medicine, Texas A&M Health Science Center, College Station, TX Emily.k.tribble@gmail.com vytas@tamhsc.edu.
authors.forename: Vytas A
authors.initials: VA
authors.lastname: Bankaitis
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000737:chemistry; D005347:Fibroblasts / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D009685:Nuclear Envelope / Q000378:metabolism*; D049294:Phospholipid Transfer Proteins / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*
chemical_list: D049294:Phospholipid Transfer Proteins; D010743:Phospholipids; C493468:Pitpn protein, mouse
keywords: cell signaling; lipids; nuclear receptors/lipid ligands; phospholipids/metabolism; phospholipids/phosphatidylinositol