title: Extending SILAC to proteomics of plant cell lines.
abstract: Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) is a widespread method for metabolic labeling of cells and tissues in quantitative proteomics; however, incomplete incorporation of the label has so far restricted its wider use in plants. Here, we argue that differential labeling by two different versions of the labeled amino acids renders SILAC fully applicable to dark-grown plant cell lines. By comparing Arabidopsis thaliana cell cultures labeled with two versions of heavy Lys (Lys-4 and Lys-8), we show that this simple modification of the SILAC protocol enables similar quantitation accuracy, precision, and reproducibility as conventional SILAC in animal cells.
authors.affiliation: Proteome Center Tuebingen, University of Tuebingen, Tuebingen, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Schütz
authors.forename: Niklas
authors.initials: N
authors.lastname: Hausmann
authors.forename: Karsten
authors.initials: K
authors.lastname: Krug
authors.forename: Rüdiger
authors.initials: R
authors.lastname: Hampp
authors.forename: Boris
authors.initials: B
authors.lastname: Macek
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism; D017360:Arabidopsis / Q000166:cytology / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D018929:Cell Culture Techniques; D002460:Cell Line; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D008239:Lysine / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D013997:Time Factors
chemical_list: D000596:Amino Acids; D029681:Arabidopsis Proteins; D008239:Lysine