title: Protein Complex Purification by Affinity Capture.
abstract: Affinity capture has become a powerful technique for consistently purifying endogenous protein complexes, facilitating biochemical and biophysical assays on otherwise inaccessible biological assemblies, and enabling broader interactomic exploration. For this procedure, cells are broken and their contents separated and extracted into a solvent, permitting access to target macromolecular complexes thus released in solution. The complexes are specifically enriched from the extract onto a solid medium coupled with an affinity reagent-usually an antibody-that recognizes the target either directly or through an appended affinity tag, allowing subsequent characterization of the complex. Here, we discuss approaches and considerations for purifying endogenous yeast protein complexes by affinity capture.
authors.affiliation: Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, New York 10065.
authors.forename: John
authors.initials: J
authors.lastname: LaCava
authors.affiliation: Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, New York 10065.
authors.forename: Javier
authors.initials: J
authors.lastname: Fernandez-Martinez
authors.affiliation: Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, New York 10065.
authors.forename: Zhanna
authors.initials: Z
authors.lastname: Hakhverdyan
authors.affiliation: Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, New York 10065.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Rout
mesh_terms: D000345:Affinity Labels / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D046911:Macromolecular Substances / Q000302:isolation & purification*; D009097:Multienzyme Complexes / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins / Q000302:isolation & purification*
chemical_list: D000345:Affinity Labels; D046911:Macromolecular Substances; D009097:Multienzyme Complexes; D029701:Saccharomyces cerevisiae Proteins