15N-Metabolic labeling for comparative plasma membrane proteomics in Arabidopsis cells

• Published in: Proteomics (Weinheim) Vol. 7; no. 5; pp. 750 - 754
• Main Authors: Lanquar, Viviane, Kuhn, Lauriane, Lelièvre, Françoise, Khafif, Mehdi, Espagne, Christelle, Bruley, Christophe, Barbier-Brygoo, Hélène, Garin, Jérôme, Thomine, Sébastien
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.03.2007; WILEY‐VCH Verlag; Wiley-VCH
• Subjects: 15N-Metabolic labeling; Analytical, structural and metabolic biochemistry; Arabidopsis; Arabidopsis - metabolism; Biological and medical sciences; Cell Membrane - metabolism; Cells, Cultured; Fundamental and applied biological sciences. Psychology; Membrane proteomics; Miscellaneous; Nitrogen - metabolism; Nitrogen Isotopes; Proteins; Proteomics - methods; Membrane proteomics; Cruciferae; Arabidopsis; Dicotyledones; Angiospermae; Plasma membrane; Proteomics; 15N-Metabolic labeling; Spermatophyta
• ISSN: 1615-9853; 1615-9861
• DOI: 10.1002/pmic.200600791

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.