title: Synthesis and biological evaluation of phosphatidylinositol phosphate affinity probes.
abstract: The synthesis of the complete family of phosphatidylinositol phosphate analogues (PIPs) from five key core intermediates A-E is described. These core compounds were obtained from myo-inositol orthoformate 1 via regioselective DIBAL-H and trimethylaluminium-mediated cleavages and a resolution-protection process using camphor acetals 10. Coupling of cores A-E with phosphoramidites 34 and 38, derived from the requisite protected lipid side chains, afforded the fully-protected PIPs. Removal of the remaining protecting groups was achieved via hydrogenolysis using palladium black or palladium hydroxide on carbon in the presence of sodium bicarbonate to afford the complete family of dipalmitoyl- and amino-PIP analogues 42, 45, 50, 51, 58, 59, 67, 68, 76, 77, 82, 83, 92, 93, 99 and 100. Investigations using affinity probes incorporating these compounds have identified novel proteins involved in the PI3K intracellular signalling network and have allowed a comprehensive proteomic analysis of phosphoinositide interacting proteins.
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Stuart J
authors.initials: SJ
authors.lastname: Conway
authors.forename: James
authors.initials: J
authors.lastname: Gardiner
authors.forename: Simon J A
authors.initials: SJ
authors.lastname: Grove
authors.forename: Melloney K
authors.initials: MK
authors.lastname: Johns
authors.forename: Ze-Yi
authors.initials: ZY
authors.lastname: Lim
authors.forename: Gavin F
authors.initials: GF
authors.lastname: Painter
authors.forename: Diane E J E
authors.initials: DE
authors.lastname: Robinson
authors.forename: Christine
authors.initials: C
authors.lastname: Schieber
authors.forename: Jan W
authors.initials: JW
authors.lastname: Thuring
authors.forename: Leon S-M
authors.initials: LS
authors.lastname: Wong
authors.forename: Meng-Xin
authors.initials: MX
authors.lastname: Yin
authors.forename: Antony W
authors.initials: AW
authors.lastname: Burgess
authors.forename: Bruno
authors.initials: B
authors.lastname: Catimel
authors.forename: Phillip T
authors.initials: PT
authors.lastname: Hawkins
authors.forename: Nicholas T
authors.initials: NT
authors.lastname: Ktistakis
authors.forename: Leonard R
authors.initials: LR
authors.lastname: Stephens
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Holmes
mesh_terms: D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000378:metabolism; D006801:Humans; D008081:Liposomes; D008958:Models, Molecular; D009943:Organophosphorus Compounds / Q000138:chemical synthesis / Q000737:chemistry; D018129:Phosphatidylinositol Phosphates / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D008081:Liposomes; D009943:Organophosphorus Compounds; D018129:Phosphatidylinositol Phosphates; D011506:Proteins; C434331:phosphoramidite