title: 15N-metabolic labeling for comparative plasma membrane proteomics in Arabidopsis cells.
abstract: An important goal for proteomic studies is the global comparison of proteomes from different genotypes, tissues, or physiological conditions. This has so far been mostly achieved by densitometric comparison of spot intensities after protein separation by 2-DE. However, the physicochemical properties of membrane proteins preclude the use of 2-DE. Here, we describe the use of in vivo labeling by the stable isotope 15N as an alternative approach for comparative membrane proteomic studies in plant cells. We confirm that 15N-metabolic labeling of proteins is possible and efficient in Arabidopsis suspension cells. Quantification of 14N versus 15N MS signals reflects the relative abundance of 14N and 15N proteins in the sample analyzed. We describe the use of 15N-metabolic labeling to perform a partial comparative analysis of Arabidopsis cells following cadmium exposure. By focusing our attention on plasma membrane proteins, we were able to confidently identify proteins showing up to 5-fold regulation compared to unexposed cells. This study provides a proof of principle that 15N-metabolic labeling is a useful technique for comparative membrane proteome studies.
authors.affiliation: Institut des Sciences du Végétal, CNRS, Gif-sur-Yvette, France.
authors.forename: Viviane
authors.initials: V
authors.lastname: Lanquar
authors.forename: Lauriane
authors.initials: L
authors.lastname: Kuhn
authors.forename: Françoise
authors.initials: F
authors.lastname: Lelièvre
authors.forename: Mehdi
authors.initials: M
authors.lastname: Khafif
authors.forename: Christelle
authors.initials: C
authors.lastname: Espagne
authors.forename: Christophe
authors.initials: C
authors.lastname: Bruley
authors.forename: Hélène
authors.initials: H
authors.lastname: Barbier-Brygoo
authors.forename: Jérôme
authors.initials: J
authors.lastname: Garin
authors.forename: Sébastien
authors.initials: S
authors.lastname: Thomine
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D009584:Nitrogen / Q000378:metabolism*; D009587:Nitrogen Isotopes; D040901:Proteomics / Q000379:methods*
chemical_list: D009587:Nitrogen Isotopes; D009584:Nitrogen