title: Toward atomic force microscopy and mass spectrometry to visualize and identify lipid rafts in plasmodesmata.
abstract: Plant cell-to-cell communication is mediated by nanopores called plasmodesmata (PDs) which are complex structures comprising plasma membrane (PM), highly packed endoplasmic reticulum and numerous membrane proteins. Although recent advances on proteomics have led to insights into mechanisms of transport, there is still an inadequate characterization of the lipidic composition of the PM where membrane proteins are inserted. It has been postulated that PDs could be formed by lipid rafts, however no structural evidence has shown to visualize and analyse their lipid components. In this perspective article, we discuss proposed experiments to characterize lipid rafts and proteins in the PDs. By using atomic force microscopy (AFM) and mass spectrometry (MS) of purified PD vesicles it is possible to determine the presence of lipid rafts, specific bound proteins and the lipidomic profile of the PD under physiological conditions and after changing transport permeability. In addition, MS can determine the stoichiometry of intact membrane proteins inserted in lipid rafts. This will give novel insights into the role of membrane proteins and lipid rafts on the PD structure.
authors.affiliation: Department of Physiology, Faculty of Biological Sciences, Pontificia Universidad Católica de Chile Santiago, Chile.
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Naulin
authors.affiliation: Department of Physiology, Faculty of Biological Sciences, Pontificia Universidad Católica de Chile Santiago, Chile.
authors.forename: Natalia A
authors.initials: NA
authors.lastname: Alveal
authors.affiliation: Department of Physiology, Faculty of Biological Sciences, Pontificia Universidad Católica de Chile Santiago, Chile.
authors.forename: Nelson P
authors.initials: NP
authors.lastname: Barrera
keywords: atomic force microscopy (AFM); lipid raft; mass spectrometry (MS); membrane proteins; plasmodesmata (PDs)