Semi-enzymatic synthesis and application of ¹³C-isotopically labelled inositol-(1,4,5)-trisphosphate.

ABSTRACT: Inositol-(1,4,5)-trisphosphate (Ins(1,4,5)P₃) is a crucial secondary messenger that controls calcium (Ca²⁺) levels inside cells, yet many questions regarding Ins(1,4,5)P₃ metabolism are challenging to address with current methods. Here, a semi-enzymatic milligram scale synthesis of isotopically labeled [¹³C₆]Ins(1,4,5)P₃ is reported which then served as a substrate to monitor the activity of mammalian type II inositol 1,4,5-trisphosphate 5-phosphatase INPP5B, using NMR spectroscopy in real time. In addition, the phosphorylation sequence catalyzed by inositol polyphosphate multikinase IPMK was confirmed using [¹³C₆]Ins(1,4,5)P₃ and 2D NMR spectroscopy. The method was subsequently applied to ch

SUBMITTER: Patharkar A 

PROVIDER: S-EPMC12301848 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature