title: Purification of PP2A holoenzymes by sequential immunoprecipitation with anti-peptide antibodies.
abstract: Understanding the multiple functions of protein phosphatase 2A (PP2A) rests on elucidating the enzymatic properties of over 50 different possible forms of the PP2A holoenzyme. We describe a procedure for highly purifying each one of these forms. This procedure is based on coexpressing in 293 cells one scaffolding A subunit, one regulatory B subunit, and one catalytic C subunit, each tagged with a different sequence, and purifying the trimeric holoenzyme by three consecutive immunoprecipitations with antibodies against the tags. In a few hours and from a small number of cells, sufficient enzyme can be purified for enzymatic studies. Purification of six different holoenzymes in parallel can easily be accomplished.
authors.affiliation: Department of Pathology, University of California at San Diego, La Jolla, CA, USA.
authors.forename: Gernot
authors.initials: G
authors.lastname: Walter
authors.forename: Jin
authors.initials: J
authors.lastname: Zhou
authors.forename: Ralf
authors.initials: R
authors.lastname: Ruediger
mesh_terms: D000906:Antibodies / Q000276:immunology*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D010455:Peptides / Q000276:immunology*; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D054648:Protein Phosphatase 2; D015203:Reproducibility of Results
chemical_list: D000906:Antibodies; D010455:Peptides; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2