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Native PAGE to study the interaction between the oncosuppressor p53 and its protein ligands
Author(s) -
Lamberti Anna,
Sgammato Roberta,
Desiderio Doriana,
Punzo Chiara,
Raimo Gennaro,
Novellino Ettore,
Carotenuto Alfonso,
Masullo Mariorosario
Publication year - 2015
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201400424
Subject(s) - mdmx , mdm2 , chemistry , dissociation (chemistry) , protein–protein interaction , signalling , biophysics , small molecule , polyacrylamide gel electrophoresis , p53 protein , microbiology and biotechnology , biochemistry , apoptosis , biology , enzyme
In the present study, we investigated a new approach for studying the interaction between p53 and MDM2/X (where MDM is murine double minute protein). The method is based on the different mobility between the interacting domains of the oncosuppressor p53 and its protein ligands MDM2/X on polyacrylamide gels under native conditions. While the two proteins MDM2/X alone were able to enter the gel, the formation of a binary complex between p53 and MDM2/X prevented the gel entry. The novel technique is reliable for determining the different affinity elicited by MDM2 or MDMX toward p53, and can be useful for analyzing the dissociation power exerted by other molecules on the p53–MDM2/X complex.