Rapid estimation of the energy charge from cell lysates using matrix-assisted laser desorption/ionization mass spectrometry

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Author(s) Steinhoff, Robert F., Krismer, Jasmin, Eyer, Klaus, Fagerer, Stephan R., Ibàñez, Alfredo, Pabst, Martin, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Rapid estimation of the energy charge from cell lysates using matrix-assisted laser desorption/ionization mass spectrometry
Subtitle Role of in-source fragmentation
Author(s) Steinhoff, Robert F.
Krismer, Jasmin
Eyer, Klaus
Fagerer, Stephan R.
Ibàñez, Alfredo
Pabst, Martin
Zenobi, Renato
Journal or Series Title Analytical Biochemistry
Volume Number 447
Issue Number 1
Start Page 107
End Page 113
ISSN 0003-2697
1096-0309
Publisher Elsevier
Publication Place San Diego
Publication Date 2014-02
Keyword(s) MALDI-MS
In-source decay
Energy charge
ATP
Metabolite analysis
Nucleoside phosphates
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.
DOI 10.1016/j.ab.2013.11.013
Additional Notes Received 11 October 2013, Revised 4 November 2013, Accepted 8 November 2013
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03430
Organisational Unit(s)
NEBIS System Number 000032007
Source Database ID SCOPUS-84890905726
WOS-000330089500018
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@article{Stnhff2014,
  author = "Steinhoff, Robert F. and Krismer, Jasmin and Eyer, Klaus and Fagerer, Stephan R. and Ib{\`{a}}{\~{n}}ez, Alfredo and Pabst, Martin and Zenobi, Renato",
  title = "{R}apid estimation of the energy charge from cell lysates using matrix-assisted laser desorption/ionization mass spectrometry: {R}ole of in-source fragmentation",
  journal = "Analytical Biochemistry",
  year = 2014,
  volume = "447",
  number = "1",
  pages = "107--113",
  month = feb,
}


E-Citations record created: Tue, 07 Jan 2014, 06:58:53 CET