title: Class III peroxidases.
abstract: Class III peroxidases are heme-containing proteins of the secretory pathway with an extremely high number of isoenzymes, indicating the tremendous and important functions of this protein family. This chapter describes fractionation of the cell in subproteomes, their separation by polyacrylamide gel electrophoresis (PAGE) and visualization of peroxidase isoenzymes by heme and specific in-gel staining procedures. Soluble and membrane-bound peroxidases were separated by differential centrifugation. Aqueous polymer two-phase partitioning and discontinuous sucrose density gradient were applied to resolve peroxidase profiles of plasma membranes and tonoplast. Peroxidase isoenzymes of subproteomes were further separated by PAGE techniques such as native isoelectric focussing (IEF), high resolution clear native electrophoresis (hrCNE), and modified sodium dodecyl sulfate (modSDS)-PAGE. These techniques were used as stand-alone method or in combination for two-dimensional PAGE.
authors.affiliation: Biocentre Klein Flottbek, University of Hamburg, Hamburg, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Lüthje
authors.forename: Claudia-Nicole
authors.initials: CN
authors.lastname: Meisrimler
authors.forename: David
authors.initials: D
authors.lastname: Hopff
authors.forename: Tim
authors.initials: T
authors.lastname: Schütze
authors.forename: Jenny
authors.initials: J
authors.lastname: Köppe
authors.forename: Katrin
authors.initials: K
authors.lastname: Heino
mesh_terms: D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D010544:Peroxidases / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012995:Solubility; D014617:Vacuoles / Q000378:metabolism; D003313:Zea mays / Q000201:enzymology*
chemical_list: D010940:Plant Proteins; D010544:Peroxidases