Homogeneous luminescent quantitation of cellular guanosine and adenosine triphosphates (GTP and ATP) using QT-Luc
Adenosine triphosphate (ATP)
Capillary electrophoresis (CE)
Guanosine triphosphate (GTP)
Immunoassay
Mass spectrometry (MS)
Time-resolved luminescence (TRL)
Journal
Analytical and bioanalytical chemistry
ISSN: 1618-2650
Titre abrégé: Anal Bioanal Chem
Pays: Germany
ID NLM: 101134327
Informations de publication
Date de publication:
Nov 2023
Nov 2023
Historique:
received:
02
08
2023
accepted:
07
09
2023
revised:
04
09
2023
medline:
26
10
2023
pubmed:
16
9
2023
entrez:
15
9
2023
Statut:
ppublish
Résumé
Guanosine triphosphate (GTP) and adenosine triphosphate (ATP) are essential nucleic acid building blocks and serve as energy molecules for a wide range of cellular reactions. Cellular GTP concentration fluctuates independently of ATP and is significantly elevated in numerous cancers, contributing to malignancy. Quantitative measurement of ATP and GTP has become increasingly important to elucidate how concentration changes regulate cell function. Liquid chromatography-coupled mass spectrometry (LC-MS) and capillary electrophoresis-coupled MS (CE-MS) are powerful methods widely used for the identification and quantification of biological metabolites. However, these methods have limitations related to specialized instrumentation and expertise, low throughput, and high costs. Here, we introduce a novel quantitative method for GTP concentration monitoring (GTP-quenching resonance energy transfer (QRET)) in homogenous cellular extracts. CE-MS analysis along with pharmacological control of cellular GTP levels shows that GTP-QRET possesses high dynamic range and accuracy. Furthermore, we combined GTP-QRET with luciferase-based ATP detection, leading to a new technology, termed QT-Luc
Identifiants
pubmed: 37714971
doi: 10.1007/s00216-023-04944-9
pii: 10.1007/s00216-023-04944-9
pmc: PMC10598090
doi:
Substances chimiques
Guanosine Triphosphate
86-01-1
Adenosine Triphosphate
8L70Q75FXE
Adenosine
K72T3FS567
Guanosine
12133JR80S
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
6689-6700Subventions
Organisme : NCI NIH HHS
ID : R01 CA255331
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM144426
Pays : United States
Organisme : NINDS NIH HHS
ID : R01 NS089815
Pays : United States
Informations de copyright
© 2023. The Author(s).
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