Details zur Publikation

Referenztyp Zeitschriften
DOI / URL Link
Titel (primär) Decimal place slope: a fast and precise method for quantifying 13C incorporation levels for detecting the metabolic activity of microbial species
Autor Jehmlich, N.; Fetzer, I.; Seifert, J.; Mattow, J.; Vogt, C.; Harms, H.; Thiede, B.; Richnow, H.H.; von Bergen, M.; Schmidt, F.;
Journal / Serie Molecular & Cellular Proteomics
Erscheinungsjahr 2010
Department ISOBIO; UMB; PROTEOM;
Band/Volume 9
Heft 6
Sprache englisch;
Abstract The metabolic incorporation of stable isotopes such as 13C or 15N into proteins has become a powerful tool for qualitative and quantitative proteome studies. We recently introduced a method that monitors heavy isotope incorporation into proteins, and presented data revealing the metabolic activity of various species in a microbial consortium using this technique. To further develop our method using an LC-MS-based approach, we present here a novel approach for calculating the incorporation level of 13C into peptides by using the information given in the decimal places of peptides masses obtained by modern high-resolution MS. In the present study, the applicability of this approach is demonstrated using Pseudomonas putida ML2 proteins uniformly labeled via the consumption of [13C6]-benzene present in the medium at concentrations of 0, 10, 25, 50, and 100 atomic %. The incorporation of 13C was calculated on the basis of several labeled peptides derived from one band on an SDS-PAGE gel. The accuracy of the calculated incorporation level depended upon the number of peptide masses included in the analysis, and it was observed that at least 100 peptide masses were required to reduce the deviation below 4 atomic %. This accuracy was comparable to calculations of incorporation based on the isotope envelope. Furthermore, this method can be extended to the calculation of the labeling efficiency for a wide range of biomolecules, including RNA and DNA. The technique will therefore allow a highly accurate determination of the carbon flux in microbial consortia with a direct approach based solely on LC-MS.
ID 10121
dauerhafte UFZ-Verlinkung https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=10121
Jehmlich, N., Fetzer, I., Seifert, J., Mattow, J., Vogt, C., Harms, H., Thiede, B., Richnow, H.H., von Bergen, M., Schmidt, F. (2010):
Decimal place slope: a fast and precise method for quantifying 13C incorporation levels for detecting the metabolic activity of microbial species
Mol. Cell. Proteomics 9 (6), 1221 - 1227