title: Analyzing the in vivo status of exogenously applied auxins: a HPLC-based method to characterize the intracellularly localized auxin transporters.
abstract: Exogenous application of biologically important molecules for plant growth promotion and/or regulation is very common both in plant research and horticulture. Plant hormones such as auxins and cytokinins are classes of compounds which are often applied exogenously. Nevertheless, plants possess a well-established machinery to regulate the active pool of exogenously applied compounds by converting them to metabolites and conjugates. Consequently, it is often very useful to know the in vivo status of applied compounds to connect them with some of the regulatory events in plant developmental processes. The in vivo status of applied compounds can be measured by incubating plants with radiolabeled compounds, followed by extraction, purification, and HPLC metabolic profiling of plant extracts. Recently we have used this method to characterize the intracellularly localized PIN protein, PIN5. Here we explain the method in detail, with a focus on general application.
authors.affiliation: Institute of Science and Technology Austria (IST Austria), Klosterneuburg, Austria.
authors.forename: Sibu
authors.initials: S
authors.lastname: Simon
authors.forename: Petr
authors.initials: P
authors.lastname: Skůpa
authors.forename: Petre I
authors.initials: PI
authors.lastname: Dobrev
authors.forename: Jan
authors.initials: J
authors.lastname: Petrášek
authors.forename: Eva
authors.initials: E
authors.lastname: Zažímalová
authors.forename: Jiří
authors.initials: J
authors.lastname: Friml
mesh_terms: D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D007210:Indoleacetic Acids / Q000302:isolation & purification / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D014026:Nicotiana / Q000166:cytology / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D007210:Indoleacetic Acids; D010940:Plant Proteins