title: Isolation of the Cell Wall.
abstract: This chapter describes a method allowing the purification of the cell wall for studying both polysaccharides and proteins. The plant primary cell wall is mainly composed of polysaccharides (90-95 % in mass) and of proteins (5-10 %). At the end of growth, specialized cells may synthesize a lignified secondary wall composed of polysaccharides (about 65 %) and lignin (about 35 %). Due to its composition, the cell wall is the cellular compartment having the highest density and this property is used for its purification. It plays critical roles during plant development and in response to environmental constraints. It is largely used in the food and textile industries as well as for the production of bioenergy. All these characteristics and uses explain why its study as a true cell compartment is of high interest. The proposed method of purification can be used for large amount of material but can also be downscaled to 500 mg of fresh material. Tools for checking the quality of the cell wall preparation, such as protein analysis and microscopy observation, are also provided.
authors.affiliation: Laboratoire de Recherche en Sciences Végétales, Université de Toulouse, CNRS, UPS, 24 chemin de Borde Rouge, Auzeville, BP 42617, 31326 Castanet, Tolosan, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Canut
authors.affiliation: Laboratoire de Recherche en Sciences Végétales, Université de Toulouse, CNRS, UPS, 24 chemin de Borde Rouge, Auzeville, BP 42617, 31326 Castanet, Tolosan, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Albenne
authors.affiliation: Laboratoire de Recherche en Sciences Végétales, Université de Toulouse, CNRS, UPS, 24 chemin de Borde Rouge, Auzeville, BP 42617, 31326 Castanet, Tolosan, France. jamet@lrsv.ups-tlse.fr.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Jamet
mesh_terms: D017360:Arabidopsis / Q000737:chemistry* / Q000254:growth & development; D058431:Brachypodium / Q000737:chemistry* / Q000254:growth & development; D002458:Cell Fractionation / Q000295:instrumentation / Q000379:methods*; D002473:Cell Wall / Q000737:chemistry* / Q000648:ultrastructure; D003470:Culture Media / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D018546:Hypocotyl / Q000737:chemistry / Q000648:ultrastructure; D008853:Microscopy; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D011134:Polysaccharides / Q000737:chemistry / Q000302:isolation & purification; D012639:Seeds / Q000737:chemistry / Q000254:growth & development; D014617:Vacuoles / Q000737:chemistry*
chemical_list: D003470:Culture Media; D010940:Plant Proteins; D011134:Polysaccharides
keywords: Cell fractionation; Cell wall protein; Plant; Polysaccharides; Proteomics; Recombinant protein production