Isolation of Plasmodesmata membranes for lipidomic and proteomic analysis
Language
EN
Chapitre d'ouvrage
This item was published in
Plasmodesmata. Methods in Molecular Biology. 2022-03, vol. 2457, p. 189-207
Springer
English Abstract
Plasmodesmata (PD) are membranous intercellular nanochannels crossing the plant cell wall to connect adjacent cells in plants. Our understanding of PD function heavily relies on the identification of their molecular ...Read more >
Plasmodesmata (PD) are membranous intercellular nanochannels crossing the plant cell wall to connect adjacent cells in plants. Our understanding of PD function heavily relies on the identification of their molecular components, these being proteins or lipids. In that regard, proteomic and lipidomic analyses of purified PD represent a crucial strategy in the field. Here we describe a simple two-step purification procedure that allows isolation of pure PD-derived membranes from Arabidopsis suspension cells suitable for "omic" approaches. The first step of this procedure consists on isolating pure cell walls containing intact PD, followed by a second step which involves an enzymatic degradation of the wall matrix to release PD membranes. The PD-enriched fraction can then serve to identify the lipid and protein composition of PD using lipidomic and proteomic approaches, which we also describe in this method article.Read less <
English Keywords
Plasmodesmata
Membrane
Cell wall
Cellulase
Suspension cell
Proteomic
Lipidomic
Mass spectrometry
Arabidopsis thaliana
ANR Project
CONTACTS MEMBRANAIRES ET LE CONTROLE DE LA COMMUNICATION INTERCELLULAIRE CHEZ LES PLANTES - ANR-18-CE13-0016
Développement d'une infrastructure française distribuée pour la métabolomique dédiée à l'innovation - ANR-11-INBS-0010
Développement d'une infrastructure française distribuée pour la métabolomique dédiée à l'innovation - ANR-11-INBS-0010