Abstract
S-Adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) are important metabolites in the one-carbon cycle that modulates cellular methylation required for proliferation and epigenetic regulation. Their concentrations, synthesis, and turnover are difficult to determine conveniently and reliably. We have developed such a method by coupling a simple and rapid purification scheme that efficiently captures both compounds, with high sensitivity, sample throughput direct infusion nanoelectrospray ultra-high-resolution Fourier transform mass spectrometry (DI-nESI-UHR-FTMS). This method is compatible with Stable Isotope-Resolved Metabolomic (SIRM) analysis of numerous other metabolites. The limits of detection for both SAM and SAH were <1 nM, and the linearity range was up to 1000 nM. The method was first illustrated for SAM/SAH analysis of mouse livers, and lung adenocarcinoma A549 cells. We then applied the method to track 13C1–CH3-Met incorporation into SAM and 13C6-glucose transformation into SAM and SAH via de novo synthesis. We further used the method to show the distinct effects on A549 and H1299 cells with treatment of anti-cancer methylseleninic acid (MSA), selenite, and selenomethionine, notably SAM depletion and increased SAM to SAH ratio by MSA, which implicates altered epigenetic regulation.
Original language | English |
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Article number | 338873 |
Journal | Analytica Chimica Acta |
Volume | 1181 |
DOIs | |
State | Published - Oct 9 2021 |
Bibliographical note
Publisher Copyright:© 2021 Elsevier B.V.
Funding
This work was supported in part by NIH grants 1U24DK097215-01A1 , P01CA163223-01A1 , 5P20GM121327 , 1P30ES026529-01A1 , and Shared Resource(s) of the University of Kentucky Markey Cancer Center P30CA177558 .
Funders | Funder number |
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National Institutes of Health (NIH) | 5P20GM121327, P01CA163223-01A1, 1U24DK097215-01A1, 1P30ES026529-01A1 |
National Institutes of Health (NIH) | |
University of Kentucky Markey Cancer Center | P30CA177558 |
University of Kentucky Markey Cancer Center |
Keywords
- DI-nESI-UHR-FTMS
- Methylseleninic acid
- S-adenosylhomocysteine
- S-adenosylmethionine
- Selenite
- Stable isotope-resolved metabolomics
ASJC Scopus subject areas
- Analytical Chemistry
- Biochemistry
- Spectroscopy
- Environmental Chemistry