Regioselective approach to phosphatidylinositol 3,5-bisphosphates: syntheses of the native phospholipid and biotinylated short-chain derivative.
A selective bis-silylation of 1D-O-TBDPS-myo-inositol leads to a 1,3,5-trisubstituted inositol, which can be advanced to the headgroup of phosphatidylinositol-3,5-bisphosphate [PI(3,5)P(2)]. A mild, regioselective method for construction of the diacylglycerol moiety containing differing fatty acid chains, including the naturally occurring lipids, was developed. Their union in the synthesis of the cell-signaling molecule PI(3,5)P(2) containing the sn-1-stearoyl and sn-2-arachidonoyl groups is described. The methodology was also used to generate dioctanoyl-PI(3,5)P(2) and a previously unreported biotin-PI(3,5)P(2) conjugate, which was coupled to neutravidin beads and used to pull down PI(3,5)P(2)-binding proteins from the cytosolic extract of adrenal neurosecretory cells. We report the specific pull-down of the PI(3,5)P(2)-binding protein svp1p, a known PI(3,5)P(2) effector involved in membrane trafficking.
Anderson
Regan J
RJ
Carbohydrate Chemistry Team, Industrial Research Limited, PO Box 31-310, Lower Hutt, New Zealand.
Osborne
Shona L
SL
Meunier
Frederic A
FA
Painter
Gavin F
GF
D000818:Animals; D019921:Biotinylation; D002417:Cattle; D019439:Chromaffin Cells; D003600:Cytosol / Q000737:chemistry; D015335:Molecular Probes / Q000138:chemical synthesis*; D018129:Phosphatidylinositol Phosphates / Q000138:chemical synthesis*; D010743:Phospholipids / Q000737:chemistry*; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification* / Q000378:metabolism
D015335:Molecular Probes; D018129:Phosphatidylinositol Phosphates; D010743:Phospholipids; D011506:Proteins; C106336:phosphatidylinositol 3,5-diphosphate