title: Determination of auxin transport parameters on the cellular level.
abstract: The accumulation of radioactively labelled compounds in cells is frequently used for the determination of activities of various transport systems located at the plasma membrane, including the system for carrier-mediated transport of plant hormone auxin. The measurements of auxin transport could be performed on the tissue level as well, but for more precise quantitative analysis of activity of individual auxin carriers the model of plant cell cultures represents an invaluable tool. Here, we describe the method for the determination of the activities of auxin influx and efflux carriers in plant cells grown in a suspension using radiolabelled synthetic auxins 2,4-dichlorophenoxyacetic acid (2,4-D) and naphthalene-1-acetic acid (NAA). By making use of specific inhibitors of active auxin influx and efflux, as well as cell lines overexpressing or silencing particular auxin carriers, this method allows the determination of kinetic parameters of auxin flow across the plasma membrane and the activity of those carriers.
authors.affiliation: Institute of Experimental Botany, ASC, Prague, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Petrášek
authors.forename: Martina
authors.initials: M
authors.lastname: Laňková
authors.forename: Eva
authors.initials: E
authors.lastname: Zažímalová
mesh_terms: D017360:Arabidopsis / Q000166:cytology / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000378:metabolism; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism; D018929:Cell Culture Techniques; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D007210:Indoleacetic Acids / Q000378:metabolism*; D007700:Kinetics; D064688:Metabolic Flux Analysis; D014026:Nicotiana / Q000166:cytology / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D007210:Indoleacetic Acids