Determination of Multimodal Isotopic Distributions: The Case of a (15)N Labeled Protein Produced into Hairy Roots

• Published in: Analytical chemistry (Washington) 2015-06, Vol.87 (12), p.5938-5946
• Main Authors: Trouillard, Romain, Hubert-Roux, Marie, Tognetti, Vincent, Guilhaudis, Laure, Plasson, Carole, Menu-Bouaouiche, Laurence, Coquet, Laurent, Guerineau, François, Hardouin, Julie, Ele Ekouna, Jean-Pierre, Cosette, Pascal, Lerouge, Patrice, Boitel-Conti, Michèle, Afonso, Carlos, Ségalas-Milazzo, Isabelle
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Brassica rapa - chemistry; Chemical Sciences; Green Fluorescent Proteins - analysis; Humans; Mass Spectrometry; Nitrogen Isotopes; Plant Roots - chemistry
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.5b01558

Isotopic labeling is widely used in various fields like proteomics, metabolomics, fluxomics, as well as in NMR structural studies, but it requires an efficient determination of the isotopic enrichment. Mass spectrometry is the method of choice for such analysis. However, when complex expression systems like hairy roots are used for production, multiple populations of labeled proteins may be obtained. If the isotopic incorporation determination is actually well-known for unimodal distributions, the multimodal distributions have scarcely been investigated. Actually, only a few approaches allow the determination of the different labeled population proportions from multimodal distributions. Furthermore, they cannot be used when the number of the populations and their respective isotope ratios are unknown. The present study implements a new strategy to measure the (15)N labeled populations inside a multimodal distribution knowing only the peptide sequence and peak intensities from mass spectrometry analyses. Noteworthy, it could be applied to other elements, like carbon and hydrogen, and extended to a larger range of biomolecules.