Difference between Extra- and Intracellular T1 Values of Carboxylic Acids Affects the Quantitative Analysis of Cellular Kinetics by Hyperpolarized NMR

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Abstract

Incomplete knowledge of the longitudinal relaxationtime constant (T1) leads to incorrect assumptions in quantita-tive kinetic models of cellular systems, studied by hyper-polarized real-time NMR. Using an assay that measures theintracellular signal of small carboxylic acids in living cells, theintracellular T1of the carboxylic acid moiety of acetate, keto-isocaproate, pyruvate, and butyrate was determined. Theintracellular T1is shown to be up to four-fold shorter thanthe extracellular T1. Such a large difference in T1valuesbetween the inside and the outside of the cell has significantinfluence on the quantification of intracellular metabolicactivity. It is expected that the significantly shorter T1valueof the carboxylic moieties inside cells is a result of macro-molecular crowding. An artificial cytosol has been preparedand applied to predict the T1of other carboxylic acids. Wedemonstrate the value of this prediction tool.
Original languageEnglish
JournalAngewandte Chemie
Volume128
Issue number43
Pages (from-to)13765–13768
Number of pages5
ISSN0044-8249
DOIs
Publication statusPublished - 2016

Keywords

  • Carbonsäuren
  • Lösungs-DNP
  • Hyperpolarisation
  • Longitudinale Relaxation
  • NMR-Spektroskopie

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