Haas, TM;
Mundinger, S;
Qiu, D;
Jork, N;
Ritter, K;
Duerr-Mayer, T;
Ripp, A;
... Jessen, HJ; + view all
(2021)
Stable Isotope Phosphate Labelling of Diverse Metabolites is Enabled by a Family of O-18-Phosphoramidites**.
Angewandte Chemie International Edition
10.1002/anie.202112457.
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Abstract
A family of 18O2-phosphoramidites facilitates synthetic access on gram-scale to various isotopically pure 18O-labelled phosphate products, like nucleotides, inositol phosphates, polyphosphates, and DNA. The utility of these 18O-natural products is underlined in the assignment of various metabolites from biological matrices using capillary electrophoresis electrospray ionisation triple quadrupole mass spectrometry. Stable isotope labelling is state-of-the-art in quantitative mass spectrometry, yet often accessing the required standards is cumbersome and very expensive. Here, a unifying synthetic concept for 18O-labelled phosphates is presented, based on a family of modified 18O2-phosphoramidite reagents. This toolbox offers access to major classes of biologically highly relevant phosphorylated metabolites as their isotopologues including nucleotides, inositol phosphates, -pyrophosphates, and inorganic polyphosphates. 18O-enrichment ratios >95 % and good yields are obtained consistently in gram-scale reactions, while enabling late-stage labelling. We demonstrate the utility of the 18O-labelled inositol phosphates and pyrophosphates by assignment of these metabolites from different biological matrices. We demonstrate that phosphate neutral loss is negligible in an analytical setup employing capillary electrophoresis electrospray ionisation triple quadrupole mass spectrometry.
| Type: | Article |
|---|---|
| Title: | Stable Isotope Phosphate Labelling of Diverse Metabolites is Enabled by a Family of O-18-Phosphoramidites** |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1002/anie.202112457 |
| Publisher version: | https://doi.org/10.1002/anie.202112457 |
| Language: | English |
| Additional information: | This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third-party material in this article are included in the Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
| Keywords: | Science & Technology, Physical Sciences, Chemistry, Multidisciplinary, Chemistry, capillary electrophoresis, mass spectrometry, nucleotides, phosphorylation, stable isotope labelling, KINASE ASSAY, MASS-SPECTROMETRY, O-18, MECHANISM, SITES |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Lab for Molecular Cell Bio MRC-UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10139654 |
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