Synthesis and biological evaluation of phosphatidylinositol phosphate affinity probes.
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.
Conway
Stuart J
SJ
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
Gardiner
James
J
Grove
Simon J A
SJ
Johns
Melloney K
MK
Lim
Ze-Yi
ZY
Painter
Gavin F
GF
Robinson
Diane E J E
DE
Schieber
Christine
C
Thuring
Jan W
JW
Wong
Leon S-M
LS
Yin
Meng-Xin
MX
Burgess
Antony W
AW
Catimel
Bruno
B
Hawkins
Phillip T
PT
Ktistakis
Nicholas T
NT
Stephens
Leonard R
LR
Holmes
Andrew B
AB
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
D008081:Liposomes; D009943:Organophosphorus Compounds; D018129:Phosphatidylinositol Phosphates; D011506:Proteins; C434331:phosphoramidite