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Journal article
LC-quadrupole/Orbitrap high-resolution mass spectrometry enables stable isotope-resolved simultaneous quantification and C-13-isotopic labeling of acyl-coenzyme A thioesters
Analytical and bioanalytical chemistry, v 408(13), pp 3651-3658
01 May 2016
PMID: 26968563
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Acyl-coenzyme A (acyl-CoA) thioesters are evolutionarily conserved, compartmentalized, and energetically activated substrates for biochemical reactions. The ubiquitous involvement of acyl-CoA thioesters in metabolism, including the tricarboxylic acid cycle, fatty acid metabolism, amino acid degradation, and cholesterol metabolism highlights the broad applicability of applied measurements of acyl-CoA thioesters. However, quantitation of acyl-CoA levels provides only one dimension of metabolic information and a more complete description of metabolism requires the relative contribution of different precursors to individual substrates and pathways. Using two distinct stable isotope labeling approaches, acyl-CoA thioesters can be labeled with either a fixed [C-13(3) N-15(1)] label derived from pantothenate into the CoA moiety or via variable [C-13] labeling into the acyl chain from metabolic precursors. Liquid chromatography-hybrid quadrupole/Orbitrap high-resolution mass spectrometry using parallel reaction monitoring, but not single ion monitoring, allowed the simultaneous quantitation of acyl-CoA thioesters by stable isotope dilution using the [C-13(3) N-15(1)] label and measurement of the incorporation of labeled carbon atoms derived from [C-13(6)]-glucose, [C-13(5) N-15(2)]-glutamine, and [C-13(3)]-propionate. As a proof of principle, we applied this method to human B cell lymphoma (WSU-DLCL2) cells in culture to precisely describe the relative pool size and enrichment of isotopic tracers into acetyl-, succinyl-, and propionyl-CoA. This method will allow highly precise, multiplexed, and stable isotope-resolved determination of metabolism to refine metabolic models, characterize novel metabolism, and test modulators of metabolic pathways involving acyl-CoA thioesters.
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Details
- Title
- LC-quadrupole/Orbitrap high-resolution mass spectrometry enables stable isotope-resolved simultaneous quantification and C-13-isotopic labeling of acyl-coenzyme A thioesters
- Creators
- Alexander J. Frey - Drexel UniversityDaniel R. Feldman - Drexel UniversitySophie Trefely - Drexel UniversityAndrew J. Worth - Bristol-Myers SquibbSankha S. Basu - Brigham and Women's HospitalNathaniel W. Snyder - Drexel University
- Publication Details
- Analytical and bioanalytical chemistry, v 408(13), pp 3651-3658
- Publisher
- Springer Nature
- Number of pages
- 8
- Grant note
- P30ES013508 / NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Environmental Health Sciences (NIEHS) Pennsylvania Department of Health Commonwealth Universal Research Enhancement (CURE) grant K22ES26235 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA R21HD087866 / EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- A.J. Drexel Autism Institute
- Web of Science ID
- WOS:000374658400027
- Scopus ID
- 2-s2.0-84960405794
- Other Identifier
- 991019167979504721
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- Collaboration types
- Industry collaboration
- Domestic collaboration
- Web of Science research areas
- Biochemical Research Methods
- Chemistry, Analytical