title: Chemical synthesis and molecular recognition of phosphatase-resistant analogues of phosphatidylinositol-3-phosphate.
abstract: The remodeling of phosphatidylinositol polyphosphates in cellular membranes by phosphatases and kinases orchestrates the signaling by these lipids in space and time. To provide chemical tools to study the changes in cell physiology mediated by these lipids, three new metabolically stabilized (ms) analogues of phosphatidylinositol-3-phosphate (PtdIns(3)P) were synthesized. We describe herein the total asymmetric synthesis of 3-methylphosphonate, 3-(monofluoromethyl)phosphonate and 3-phosphorothioate analogues of PtdIns(3)P. From differentially protected D-myo-inositol key intermediates, a versatile phosphoramidite reagent was employed in the synthesis of PtdIns(3)P analogues with diacylglyceryl moieties containing dioleoyl, dipalmitoyl, and dibutyryl chains. In addition, we introduce a new phosphorylation reagent, (monofluoromethyl)phosphonyl chloride, which has general applications for the preparation of "pKa-matched" monofluorophosphonates. These ms-PtdIns(3)P analogues exhibited reduced binding activities with 15N-labeled FYVE and PX domains, as significant 1H and 15N chemical shift changes in the FYVE domain were induced by titrating ms-PtdIns(3)P analogues into membrane-mimetic dodecylphosphocholine micelles. In addition, the PtdIns(3)P analogues with dioleoyl and dipalmitoyl chains were substrates for the 5-kinase enzyme PIKfyve; the corresponding phosphorylated ms-PI(3,5)P2 products were detected by radio-TLC analysis.
authors.affiliation: Department of Medicinal Chemistry, The University of Utah, 419 Wakara Way, Suite 205, Salt Lake City, Utah 84108-1257, USA.
authors.forename: Yong
authors.initials: Y
authors.lastname: Xu
authors.forename: Stephanie A
authors.initials: SA
authors.lastname: Lee
authors.forename: Tatiana G
authors.initials: TG
authors.lastname: Kutateladze
authors.forename: Diego
authors.initials: D
authors.lastname: Sbrissa
authors.forename: Assia
authors.initials: A
authors.lastname: Shisheva
authors.forename: Glenn D
authors.initials: GD
authors.lastname: Prestwich
mesh_terms: D006801:Humans; D009943:Organophosphorus Compounds / Q000138:chemical synthesis / Q000737:chemistry; D063086:Organothiophosphates / Q000138:chemical synthesis* / Q000378:metabolism; D018129:Phosphatidylinositol Phosphates / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010766:Phosphorylation; D017434:Protein Structure, Tertiary
chemical_list: D009943:Organophosphorus Compounds; D063086:Organothiophosphates; D018129:Phosphatidylinositol Phosphates; C032627:methylphosphonic acid; D010744:Phosphoric Monoester Hydrolases