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- W630560838 abstract "Quantitative analysis by mass spectrometry (MS) is a major challenge inproteomics as the correlation between analyte concentration and signalintensity is often poor due to varying ionisation efficiencies in thepresence of molecular competitors. However, relative quantitationmethods that utilise differential stable isotope labelling and massspectrometric detection are available. Many drawbacks inherent tochemical labelling methods (ICAT, iTRAQ) can be overcome bymetabolic labelling with amino acids containing stable isotopes (e.g. 13Cand/or 15N) in methods such as Stable Isotope Labelling with Aminoacids in Cell culture (SILAC). SILAC has also been used for labelling ofproteins in plant cell cultures (1) but is not suitable for whole plantlabelling. Plants are usually autotrophic (fixing carbon from atmosphericCO2) and, thus, labelling with carbon isotopes becomes impractical. Inaddition, SILAC is expensive.Recently, Arabidopsis cell cultures were labelled with 15N in a mediumcontaining nitrate as sole nitrogen source. This was shown to be suitablefor quantifying proteins and nitrogen-containing metabolites from this cellculture (2,3).Labelling whole plants, however, offers the advantage of studyingquantitatively the response to stimulation or disease of a whole multicellularorganism or multi-organism systems at the molecular level.Furthermore, plant metabolism enables the use of inexpensive labellingmedia without introducing additional stress to the organism. And finally,hydroponics is ideal to undertake metabolic labelling under extremelywell-controlled conditions.We demonstrate the suitability of metabolic 15N hydroponic isotopelabelling of entire plants (HILEP) for relative quantitative proteomicanalysis by mass spectrometry. To evaluate this methodology,Arabidopsis plants were grown hydroponically in 14N and 15N media and subjected to oxidative stress." @default.
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- W630560838 date "2009-06-07" @default.
- W630560838 modified "2023-10-11" @default.
- W630560838 title "Hydroponic isotope labeling of entire plants (HILEP) for quantitative plant proteomics" @default.
- W630560838 hasPublicationYear "2009" @default.
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