Jump to main content
Jump to site search

Issue 20, 2019
Previous Article Next Article

Harnessing 13C-labeled myo-inositol to interrogate inositol phosphate messengers by NMR

Author affiliations

Abstract

Inositol poly- and pyrophosphates (InsPs and PP-InsPs) are an important group of metabolites and mediate a wide range of processes in eukaryotic cells. To elucidate the functions of these molecules, robust techniques for the characterization of inositol phosphate metabolism are required, both at the biochemical and the cellular level. Here, a new tool-set is reported, which employs uniformly 13C-labeled compounds ([13C6]myo-inositol, [13C6]InsP5, [13C6]InsP6, and [13C6]5PP-InsP5), in combination with commonly accessible NMR technology. This approach permitted the detection and quantification of InsPs and PP-InsPs within complex mixtures and at physiological concentrations. Specifically, the enzymatic activity of IP6K1 could be monitored in vitro in real time. Metabolic labeling of mammalian cells with [13C6]myo-inositol enabled the analysis of cellular pools of InsPs and PP-InsPs, and uncovered high concentrations of 5PP-InsP5 in HCT116 cells, especially in response to genetic and pharmacological perturbation. The reported method greatly facilitates the analysis of this otherwise spectroscopically silent group of molecules, and holds great promise to comprehensively analyze inositol-based signaling molecules under normal and pathological conditions.

Graphical abstract: Harnessing 13C-labeled myo-inositol to interrogate inositol phosphate messengers by NMR

Back to tab navigation

Supplementary files

Publication details

The article was received on 10 Jan 2019, accepted on 17 Apr 2019 and first published on 23 Apr 2019


Article type: Edge Article
DOI: 10.1039/C9SC00151D
Chem. Sci., 2019,10, 5267-5274
  • Open access: Creative Commons BY-NC license
  •   Request permissions

    Harnessing 13C-labeled myo-inositol to interrogate inositol phosphate messengers by NMR

    R. K. Harmel, R. Puschmann, M. Nguyen Trung, A. Saiardi, P. Schmieder and D. Fiedler, Chem. Sci., 2019, 10, 5267
    DOI: 10.1039/C9SC00151D

    This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence. Material from this article can be used in other publications provided that the correct acknowledgement is given with the reproduced material and it is not used for commercial purposes.

    Reproduced material should be attributed as follows:

    • For reproduction of material from NJC:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the Centre National de la Recherche Scientifique (CNRS) and the RSC.
    • For reproduction of material from PCCP:
      [Original citation] - Published by the PCCP Owner Societies.
    • For reproduction of material from PPS:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the European Society for Photobiology, the European Photochemistry Association, and RSC.
    • For reproduction of material from all other RSC journals:
      [Original citation] - Published by The Royal Society of Chemistry.

    Information about reproducing material from RSC articles with different licences is available on our Permission Requests page.

Search articles by author

Spotlight

Advertisements