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Published on: March 18, 2011
Revisiting Plant Plasma Membrane Lipids in Tobacco: A Focus on Sphingolipids
Jean-Luc Cacas1, Corinne Buré1, Kevin Grosjean1
1Laboratoire de Biogenèse Membranaire, Centre National de la Recherche Scientifique-University of Bordeaux, Unité Mixte de Recherche 5200, F-33883 Villenave d'Ornon cedex, France (J.-L.C., Cl.B., J.G., F.F., L.F., V.G., E.B., S.M.);Chimie Biologie des Membranes et Nanoobjets, Unité Mixte de Recherche 5248, Centre de Génomique Fonctionnelle, Université de Bordeaux, F-33076 Bordeaux cedex, France (Co.B., J.-M.S.);Université de Bourgogne, Unité Mixte de Recherche 1347 Agroécologie, Equipes de Recherche Labellisée 6300 Centre National de la Recherche Scientifique, F-21065 Dijon cedex, France (J.-L.C., K.G., P.G.-P.);Institut National de la Recherche Agronomique, Unité Mixte de Recherche 1347 Agroécologie, Equipes de Recherche Labellisée 6300 Centre National de la Recherche Scientifique, F-21065 Dijon cedex, France (J.L., F.R., F.S.-P.);Université de Lille 1, Unité de Glycobiologie Structurale et Fonctionnelle, F-59655 Villeneuve d'Ascq, France (Y.R., E.M.);Centre National de la Recherche Scientifique, Unité Mixte de Recherche 8576, F-59655 Villeneuve d'Ascq, France (Y.R., E.M.);Laboratoire de Biophysique Moléculaire aux Interfaces, Université de Liège, B-5030 Gembloux, Belgium (Cl.B., M.D., L.L.); andInstitut des Sciences de la Vigne et du Vin, Groupe d'Etude des Substances Végétales à Activité Biologique, University of Bordeaux, Equipe Associée 3675, F-33400 Talence, France (S.C.).
Abstract:
The lipid composition of plasma membrane (PM) and the corresponding detergent-insoluble membrane (DIM) fraction were analyzed with a specific focus on highly polar sphingolipids, so-called glycosyl inositol phosphorylceramides (GIPCs). Using tobacco (Nicotiana tabacum) 'Bright Yellow 2' cell suspension and leaves, evidence is provided that GIPCs represent up to 40 mol % of the PM lipids. Comparative analysis of DIMs with the PM showed an enrichment of 2-hydroxylated very-long-chain fatty acid-containing GIPCs and polyglycosylated GIPCs in the DIMs. Purified antibodies raised against these GIPCs were further used for immunogold-electron microscopy strategy, revealing the distribution of polyglycosylated GIPCs in domains of 35 ± 7 nm in the plane of the PM. Biophysical studies also showed strong interactions between GIPCs and sterols and suggested a role for very-long-chain fatty acids in the interdigitation between the two PM-composing monolayers. The ins and outs of lipid asymmetry, raft formation, and interdigitation in plant membrane biology are finally discussed.
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