Stable Isotopomers
of myo-Inositol
Uncover a Complex MINPP1-Dependent Inositol Phosphate Network
Posted on 2022-12-05 - 18:07
The water-soluble
inositol phosphates (InsPs) represent
a functionally
diverse group of small-molecule messengers involved in a myriad of
cellular processes. Despite their centrality, our understanding of
human InsP metabolism is incomplete because the available analytical
toolset to characterize and quantify InsPs in complex samples is limited.
Here, we have synthesized and applied symmetrically and unsymmetrically 13C-labeled myo-inositol and inositol phosphates.
These probes were utilized in combination with nuclear magnetic resonance
spectroscopy (NMR) and capillary electrophoresis mass spectrometry
(CE-MS) to investigate InsP metabolism in human cells. The labeling
strategy provided detailed structural information via NMRdown
to individual enantiomerswhich overcomes a crucial blind spot
in the analysis of InsPs. We uncovered a novel branch of InsP dephosphorylation
in human cells which is dependent on MINPP1, a phytase-like enzyme
contributing to cellular homeostasis. Detailed characterization of
MINPP1 activity in vitro and in cells showcased the unique reactivity
of this phosphatase. Our results demonstrate that metabolic labeling
with stable isotopomers in conjunction with NMR spectroscopy and CE-MS
constitutes a powerful tool to annotate InsP networks in a variety
of biological contexts.
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Nguyen Trung, Minh; Kieninger, Stefanie; Fandi, Zeinab; Qiu, Danye; Liu, Guizhen; Mehendale, Neelay K.; et al. (2022). Stable Isotopomers
of myo-Inositol
Uncover a Complex MINPP1-Dependent Inositol Phosphate Network. ACS Publications. Collection. https://doi.org/10.1021/acscentsci.2c01032
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AUTHORS (10)
MN
Minh Nguyen Trung
SK
Stefanie Kieninger
ZF
Zeinab Fandi
DQ
Danye Qiu
GL
Guizhen Liu
NM
Neelay K. Mehendale
AS
Adolfo Saiardi
HJ
Henning Jessen
BK
Bettina Keller
DF
Dorothea Fiedler
KEYWORDS
molecule messengers involvedlike enzyme contributingindividual enantiomers functionally diverse groupcrucial blind spotavailable analytical toolsetinvestigate insp metabolismannotate insp networkshuman insp metabolismsoluble inositol phosphates13 supinsp dephosphorylationinositol phosphatesmyo human cellsinositol uncover>- inositolunique reactivitystable isotopomersresults demonstratepowerful toolnovel branchmyo -metabolic labelingdetailed characterizationcomplex samplescellular processescellular homeostasiscells showcasedbiological contextsapplied symmetrically