title: Single-step Strep-tag purification for the isolation and identification of protein complexes from mammalian cells.
abstract: Identification of protein complexes is the key to understanding cellular functions. In this study, we present a novel method for the identification of multiprotein complexes from mammalian cells. By using the Strep-tag affinity chromatography method, enabling fast and simple one-step purification, coupled with competitive elution under physiological conditions, we successfully purified a PP2A holoenzyme protein complex from a cultured mammalian cancer cell line. We identified, by mass spectrometry, both known and novel interacting proteins for PP2A, and demonstrate that the purified PP2A complex is functional. The benefits and potential applications of the Strep-tag method for protein complex purification are discussed.
authors.affiliation: Centre for Biotechnology, University of Turku and Abo Akademi University, Turku, Finland.
authors.forename: Melissa R
authors.initials: MR
authors.lastname: Junttila
authors.forename: Susanna
authors.initials: S
authors.lastname: Saarinen
authors.forename: Thomas
authors.initials: T
authors.lastname: Schmidt
authors.forename: Juergen
authors.initials: J
authors.lastname: Kast
authors.forename: Jukka
authors.initials: J
authors.lastname: Westermarck
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D046912:Multiprotein Complexes; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D010749:Phosphoprotein Phosphatases