1 research outputs found
Profiling Reactive Metabolites via Chemical Trapping and Targeted Mass Spectrometry
Metabolomic profiling studies aim
to provide a comprehensive, quantitative,
and dynamic portrait of the endogenous metabolites in a biological
system. While contemporary technologies permit routine profiling of
many metabolites, intrinsically labile metabolites are often improperly
measured or omitted from studies due to unwanted chemical transformations
that occur during sample preparation or mass spectrometric analysis.
The primary glycolytic metabolite 1,3-bisphosphoglyceric acid (1,3-BPG)
typifies this class of metabolites, and, despite its central position
in metabolism, has largely eluded analysis in profiling studies. Here
we take advantage of the reactive acylphosphate group in 1,3-BPG to
chemically trap the metabolite with hydroxylamine during metabolite
isolation, enabling quantitative analysis by targeted LC–MS/MS.
This approach is compatible with complex cellular metabolome, permits
specific detection of the reactive (1,3-) instead of nonreactive (2,3-)
BPG isomer, and has enabled direct analysis of dynamic 1,3-BPG levels
resulting from perturbations to glucose processing. These studies
confirmed that standard metabolomic methods misrepresent cellular
1,3-BPG levels in response to altered glucose metabolism and underscore
the potential for chemical trapping to be used for other classes of
reactive metabolites