title: Isolation of plasmodesmata from Arabidopsis suspension culture cells.
abstract: Due to their position firmly anchored within the plant cell wall, plasmodesmata (PD) are notoriously difficult to isolate from plant tissue. Yet, getting access to isolated PD represents the most straightforward strategy for the identification of their molecular components. Proteomic and lipidomic analyses of such PD fractions have provided and will continue to provide critical information on the functional and structural elements that define these membranous nano-pores. Here, we describe a two-step simple purification procedure that allows isolation of pure PD-derived membranes from Arabidopsis suspension cells. The first step of this procedure consists in isolating cell wall fragments containing intact PD while free of contamination from other cellular compartments. The second step relies on an enzymatic degradation of the wall matrix and the subsequent release of "free" PD. Isolated PD membranes provide a suitable starting material for the analysis of PD-associated proteins and lipids.
authors.affiliation: Laboratory of Membrane Biogenesis, CNRS UMR5200, University of Bordeaux, Campus INRA de Bordeaux, 71 avenue E. Bourlaux, 33883, Villenave d'Ornon Cedex, France.
authors.forename: Magali S
authors.initials: MS
authors.lastname: Grison
authors.forename: Lourdes
authors.initials: L
authors.lastname: Fernandez-Calvino
authors.forename: Sébastien
authors.initials: S
authors.lastname: Mongrand
authors.forename: Emmanuelle M F
authors.initials: EM
authors.lastname: Bayer
mesh_terms: D017360:Arabidopsis / Q000737:chemistry* / Q000166:cytology; D029681:Arabidopsis Proteins / Q000235:genetics; D015415:Biomarkers / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics; D018929:Cell Culture Techniques; D002458:Cell Fractionation / Q000379:methods*; D002473:Cell Wall / Q000737:chemistry*; D044602:Cellulases / Q000737:chemistry; D015870:Gene Expression; D007150:Immunohistochemistry; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins / Q000235:genetics; D008853:Microscopy; D031425:Plasmodesmata / Q000378:metabolism / Q000648:ultrastructure*
chemical_list: D029681:Arabidopsis Proteins; D015415:Biomarkers; D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; C542004:PDCB1 protein, Arabidopsis; C528450:PDLP1a protein, Arabidopsis; D044602:Cellulases