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Molecular & Cellular Proteomics 8:2186-2198, 2009.
© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Quantitative Proteomics Reveals a Dynamic Association of Proteins to Detergent-resistant Membranes upon Elicitor Signaling in Tobacco^*,

Thomas Stanislas^,, David Bouyssie^,¶,||, Michel Rossignol^¶,||,**, Simona Vesa, Jérôme Fromentin, Johanne Morel, Carole Pichereaux^¶,||,**, Bernard Monsarrat^¶,||, and Françoise Simon-Plas^,

From the Institut National de la Recherche Agronomique (INRA), Unité Mixte de Recherche (UMR) Plante Microbe Environnement 1088/CNRS 5184/Université de Bourgogne, 17 Rue Sully, BP 86510 F-21000 Dijon, France,
¶Institut de Pharmacologie et de Biologie Structurale (IPBS), CNRS, 205 route de Narbonne, F-31077 Toulouse, France,
||IPBS, Université Paul Sabatier, Université de Toulouse, F-31077 Toulouse, France, and
**IPBS, Institut Fédératif de Recherche 40 Plateforme Protéomique, 205 route de Narbonne, F-31077 Toulouse, France

A large body of evidence from the past decade supports the existence, in membrane from animal and yeast cells, of functional microdomains playing important roles in protein sorting, signal transduction, or infection by pathogens. In plants, as previously observed for animal microdomains, detergent-resistant fractions, enriched in sphingolipids and sterols, were isolated from plasma membrane. A characterization of their proteic content revealed their enrichment in proteins involved in signaling and response to biotic and abiotic stress and cell trafficking suggesting that these domains were likely to be involved in such physiological processes. In the present study, we used ¹⁴N/¹⁵N metabolic labeling to compare, using a global quantitative proteomics approach, the content of tobacco detergent-resistant membranes extracted from cells treated or not with cryptogein, an elicitor of defense reaction. To analyze the data, we developed a software allowing an automatic quantification of the proteins identified. The results obtained indicate that, although the association to detergent-resistant membranes of most proteins remained unchanged upon cryptogein treatment, five proteins had their relative abundance modified. Four proteins related to cell trafficking (four dynamins) were less abundant in the detergent-resistant membrane fraction after cryptogein treatment, whereas one signaling protein (a 14-3-3 protein) was enriched. This analysis indicates that plant microdomains could, like their animal counterpart, play a role in the early signaling process underlying the setup of defense reaction. Furthermore proteins identified as differentially associated to tobacco detergent-resistant membranes after cryptogein challenge are involved in signaling and vesicular trafficking as already observed in similar studies performed in animal cells upon biological stimuli. This suggests that the ways by which the dynamic association of proteins to microdomains could participate in the regulation of the signaling process may be conserved between plant and animals.

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