title: Interaction of p53 with cellular proteins.
abstract: Cellular proteins that interact with p53 play a major role in both positive and negative regulation of this tumor suppressor and can fine-tune its response to specific cellular stresses. As a consequence, p53 biology will not be complete until these interacting proteins are fully characterized. This chapter outlines two methods for identifying and characterizing p53-binding proteins: (i) glutathione-S-transferase (GST) protein-protein interaction assay; and (ii) co-immunoprecipitation (co-IP) assay. These two methods are ideal for any laboratory to perform; the assays are short in duration, do not require specialized expertise to establish in the laboratory, give reliable and reproducible data, and are cost-efficient, because few reagents are needed. This chapter gives a basic description of these two techniques and provides tips that are not found in other protocol manuals on how to achieve the best results.
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health and Science University, Portland, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Keller
authors.forename: Shelya X
authors.initials: SX
authors.lastname: Zeng
authors.forename: Hua
authors.initials: H
authors.lastname: Lu
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D051379:Mice; D011233:Precipitin Tests / Q000379:methods*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; D016159:Tumor Suppressor Protein p53; D005982:Glutathione Transferase