Development of isotope-enriched phosphatidylinositol-4- and 5-phosphate cellular mass spectrometry probes

• Published in: Chemical science (Cambridge) 2021-02, Vol.12 (7), p.2549-2557
• Main Authors: Joffrin, Amélie M, Saunders, Alex M, Barneda, David, Flemington, Vikki, Thompson, Amber L, Sanganee, Hitesh J, Conway, Stuart J
• Format: Article
• Language: English
• Subjects: Crystallography; Deuteration; Enantiomers; Mass spectrometry; NMR; Nuclear magnetic resonance; Phospholipids; Synthesis
• ISSN: 2041-6520; 2041-6539
• DOI: 10.1039/d0sc06219g

Synthetic phosphatidylinositol phosphate (PtdIns P n ) derivatives play a pivotal role in broadening our understanding of PtdIns P n metabolism. However, the development of such tools is reliant on efficient enantioselective and regioselective synthetic strategies. Here we report the development of a divergent synthetic route applicable to the synthesis of deuterated PtdIns4 P and PtdIns5 P derivatives. The synthetic strategy developed involves a key enzymatic desymmetrisation step using Lipozyme TL-IM®. In addition, we optimised the large-scale synthesis of deuterated myo -inositol, allowing for the preparation of a series of saturated and unsaturated deuterated PtdIns4 P and PtdIns5 P derivatives. Experiments in MCF7 cells demonstrated that these deuterated probes enable quantification of the corresponding endogenous phospholipids in a cellular setting. Overall, these deuterated probes will be powerful tools to help improve our understanding of the role played by PtdIns P n in physiology and disease. We report the synthesis of deuterium-labelled derivatives of phosphatidylinositol 4-phosphate and phosphatidylinositol 5-phosphate, and demonstrate their use in quantifying levels of endogenous phospholipids in cells.