Chemical Synthesis and Molecular Recognition of Phosphatase-Resistant Analogues of Phosphatidylinositol-3-phosphate

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• 作者：Yong Xu ; Stephanie A. Lee ; Tatiana G. Kutateladze ; Diego Sbrissa ; Assia Shisheva ; and Glenn D. Prestwich
• 刊名：Journal of the American Chemical Society
• 出版年：2006
• 出版时间：January 25, 2006
• 年：2006
• 卷：128
• 期：3
• 页码：885 - 897
• 全文大小：386K
• 年卷期：v.128,no.3(January 25, 2006)
• ISSN：1520-5126

文摘

The remodeling of phosphatidylinositol polyphosphates in cellular membranes by phosphatasesand kinases orchestrates the signaling by these lipids in space and time. To provide chemical tools tostudy 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 totalasymmetric synthesis of 3-methylphosphonate, 3-(monofluoromethyl)phosphonate and 3-phosphorothioateanalogues of PtdIns(3)P. From differentially protected D-myo-inositol key intermediates, a versatilephosphoramidite reagent was employed in the synthesis of PtdIns(3)P analogues with diacylglyceryl moietiescontaining dioleoyl, dipalmitoyl, and dibutyryl chains. In addition, we introduce a new phosphorylation reagent,(monofluoromethyl)phosphonyl chloride, which has general applications for the preparation of "pK_a-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 domainwere 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-kinaseenzyme PIKfyve; the corresponding phosphorylated ms-PI(3,5)P₂ products were detected by radio-TLCanalysis.