A DIGE-based approach to study interacting proteins.
A full spectrum of high-throughput protein identification and characterization approaches has been developed for protein profiling. However, the most demanding field to better understanding protein interactions known as the "interactome" is still of a perpetual need for modern proteomics. Recently developed DIGE (difference in-gel electrophoresis) system may be of potential use when studying interacting proteins. In this work we applied DIGE technique on native gel electrophoresis to study protein-protein interactions. As a proof of principle, we utilized an in vitro interaction model between p53 and HDM2 proteins. In parallel, we also showed interaction of these proteins using fluorescently labelled p53- or HDM2-immunoprecipitation pellets. Thus, we believe this study shows a good potential for investigating various interacting partners and benefits towards creation of interactome.
Lyakhovich
Alex
A
Group of Mutagenesis, Department of Genetics and Microbiology, Universitat, Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain. alex.lyakhovich@uab.es
Canals
Francesc
F
Nosov
Michael
M
Surralles
Jordi
J
D004586:Electrophoresis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D051736:Proto-Oncogene Proteins c-mdm2 / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000302:isolation & purification; D015203:Reproducibility of Results; D016159:Tumor Suppressor Protein p53 / Q000302:isolation & purification
D011506:Proteins; D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2