title: The detection and quantitation of protein oligomerization.
abstract: There are many different techniques available to biologists and biochemists that can be used to detect and characterize the self-association of proteins. Each technique has strengths and weaknesses and it is often useful to combine several approaches to maximize the former and minimize the latter. Here we review a range of methodologies that identify protein self-association and/or allow the stoichiometry and affinity of the interaction to be determined, placing an emphasis on what type of information can be obtained and outlining the advantages and disadvantages involved. In general, in vitro biophysical techniques, such as size exclusion chromatography, analytical ultracentrifugation, scattering techniques, NMR spectroscopy, isothermal titration calorimetry, fluorescence anisotropy and mass spectrometry, provide information on stoichiometry and/or binding affinities. Other approaches such as cross-linking, fluorescence methods (e.g., fluorescence correlation spectroscopy, FCS; Förster resonance energy transfer, FRET; fluorescence recovery after photobleaching, FRAP; and proximity imaging, PRIM) and complementation approaches (e.g., yeast two hybrid assays and bimolecular fluorescence complementation, BiFC) can be used to detect protein self-association in a cellular context.
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: David A
authors.initials: DA
authors.lastname: Gell
authors.forename: Richard P
authors.initials: RP
authors.lastname: Grant
authors.forename: Joel P
authors.initials: JP
authors.lastname: Mackay
mesh_terms: D002151:Calorimetry; D002850:Chromatography, Gel; D031541:Fluorescence Resonance Energy Transfer; D008027:Light; D013058:Mass Spectrometry; D057054:Molecular Imaging; D019906:Nuclear Magnetic Resonance, Biomolecular; D055503:Protein Multimerization; D021122:Protein Subunits / Q000737:chemistry* / Q000378:metabolism; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D053838:Scattering, Small Angle; D013050:Spectrometry, Fluorescence; D014461:Ultracentrifugation
chemical_list: D021122:Protein Subunits; D011506:Proteins