Abstract
The relative quantification of proteins is one of the major techniques to elucidate physiological reactions. In order to avoid artifacts due to chemical labeling the metabolic introduction of heavy isotopes into proteins and peptides is the preferred method for relative quantification. For eukaryotic cells stable isotope labeling by amino acids in cell culture (SILAC) has become the gold standard which can be readily applied to a vast number of scenarios. In the microbial realm with its highly versatile metabolic capabilities SILAC is often not feasible and using other (13)C or (15)N labeled substrates may not be practical. Here, the incorporation of heavy sulfur isotopes is a useful alternative. We therefore introduce (34)sulfur stable isotope labeling of amino acids for quantification (SULAQ34) and the corresponding tools required for spectra extraction and disintegration of the isotopic overlaps caused by the small mass shift. As a proof of principle we investigated the proteomic changes related to naphthalene degradation in P. fluorescens ATCC 17483 and uncovered a specific oxidative stress like response.
Original language | English |
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Journal | Molecular and Cellular Proteomics |
Volume | 12 |
Issue number | 8 |
Pages (from-to) | 2060-2069 |
Number of pages | 10 |
ISSN | 1535-9476 |
DOIs | |
Publication status | Published - 2013 |