title: Cell fractionation.
abstract: Proteins within a cell are localized into specific cellular compartments, allowing compartmentalization of distinct tasks. If we consider lipid bilayers as compartments, then gram-negative bacteria such as Pseudomonas aeruginosa can target proteins to five distinct locations: the cytoplasm, the inner membrane, the periplasm, the outer membrane, and the extracellular environment. In this chapter, we describe how the different compartments can be selectively isolated by a combination of centrifugation and disruption techniques. Fractionation of the cells into subcellular compartments enables protein enrichment and is essential to accurately determine the localization of specific proteins, which is the first step towards understanding the function of a protein in the cell.
authors.affiliation: Laboratoire d'Ingénierie des Systèmes Macromoléculaires, UMR7255 - CNRS - Aix Marseille Université, IMM 31, Ch. J. Aiguier, 13402, Marseille, CEDEX 20, France, bize@imm.cnrs.fr.
authors.forename: Bérengère
authors.initials: B
authors.lastname: Ize
authors.forename: Véronique
authors.initials: V
authors.lastname: Viarre
authors.forename: Romé
authors.initials: R
authors.lastname: Voulhoux
mesh_terms: D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D003470:Culture Media; D003593:Cytoplasm / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000166:cytology*; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D003470:Culture Media