Rapid estimation of the energy charge from cell lysates using matrix-assisted laser desorption/ionization mass spectrometry: Role of in-source fragmentation

Robert F. Steinhoff, Jasmin Krismer, Klaus Eyer, Stephan R. Fagerer, Alfredo Ibàñez, Martin Pabst, Renato Zenobi

Research output: Contribution to journalArticleScientificpeer-review

4 Citations (Scopus)

Abstract

Nucleotides are key players in the central energy metabolism of cells. Here we show how to estimate the energy charge from cell lysates by direct negative ion matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) using 9-aminoacridine as matrix. We found a high level of in-source decay of all the phosphorylated nucleotides, with some of them producing considerable amounts of adenosine-5′-diphosphate (ADP) fragment ions. We investigated the behavior of adenosine-5′-monophosphate (AMP), ADP, and adenosine-5′-triphosphate (ATP) as well as the cofactors coenzyme A (CoA) and acetyl-coenzyme A (ACoA) and nicotinamide adenine dinucleotides (NAD + and NADH) in detail. In-source decay of these compounds depends strongly on the applied laser power and on the extraction pulse delay. At standard instrument settings, the 9-aminoacridine (9-AA) matrix resulted in a much higher in-source decay compared with 2,4,6-trihydroxyacetophenone (2,4,6-THAP). By adding 13C-labeled ATP to a cell lysate, we were able to determine the degree of in-source decay during an experiment. Analyzing a cell extract of the monocytic cell line THP-1 with [13C]ATP as internal standard, we were able to obtain values for the energy charge that were similar to those determined by a reference liquid chromatography electrospray ionization coupled to mass spectrometry (LC-ESI-MS) method.

Original languageEnglish
Pages (from-to)107-113
JournalAnalytical Biochemistry
Volume447
Issue number1
DOIs
Publication statusPublished - 2014
Externally publishedYes

Keywords

  • ATP
  • Energy charge
  • In-source decay
  • MALDI-MS
  • Metabolite analysis
  • Nucleoside phosphates

Fingerprint

Dive into the research topics of 'Rapid estimation of the energy charge from cell lysates using matrix-assisted laser desorption/ionization mass spectrometry: Role of in-source fragmentation'. Together they form a unique fingerprint.

Cite this