title: Methods for the determination of the mass of nuclear PtdIns4P, PtdIns5P, and PtdIns(4,5)P2.
abstract: Phosphatidylinositol (PtdIns) and its phosphorylated derivatives represent less than 5% of total membrane phospholipids in cells. Despite their low abundance, they form a dynamic signaling system that is regulated in response to a variety of extra- and intracellular cues. Protein domains including PH, FYVE, ENTH, PHOX, PHD fingers, and lysine-/arginine-rich patches can bind to specific phosphoinositide isomers, which, in turn, can induce changes in the subcellular localization, posttranslational modification, protein interaction partners, or activity of the protein containing such a domain. Phosphoinositides and the enzymes that synthesize them are found in many different subcellular compartments including the nuclear matrix, heterochromatin, and sites of active RNA splicing, suggesting that phosphoinositides may regulate specific functions within the nuclear compartment. The existence of distinct subcellular pools has led to the challenging task of the quantitation of temporal and spatial changes in phosphoinositides. We report methods to measure the mass levels of three different phosphoinositides within the nuclear compartment.
authors.affiliation: Division of Cellular Biochemistry, The Netherlands Cancer Institute, Amsterdam.
authors.forename: David R
authors.initials: DR
authors.lastname: Jones
authors.forename: Yvette
authors.initials: Y
authors.lastname: Bultsma
authors.forename: Willem Jan
authors.initials: WJ
authors.lastname: Keune
authors.forename: Nullin
authors.initials: N
authors.lastname: Divecha
mesh_terms: D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry*; D005898:Glass / Q000737:chemistry; D006801:Humans; D009355:Neomycin / Q000737:chemistry; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification; D018129:Phosphatidylinositol Phosphates / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; D009355:Neomycin