MDM2-MDM4 molecular interaction investigated by atomic force spectroscopy and surface plasmon resonance.
Murine double minute 2 (MDM2) and 4 (MDM4) are known as the main negative regulators of p53, a tumor suppressor. They are able to form heterodimers that are much more effective in the downregulation of p53. Therefore, the MDM2-MDM4 complex could be a target for promising therapeutic restoration of p53 function. To this aim, a deeper understanding of the molecular mechanisms underlining the heterodimerization is needed. The kinetic and thermodynamic characterization of the MDM2-MDM4 complex was performed with two complementary approaches: atomic force spectroscopy and surface plasmon resonance. Both techniques revealed an equilibrium dissociation constant (KD ) in the micromolar range for the MDM2-MDM4 heterodimer, similar to related complexes involved in the p53 network. Furthermore, the MDM2-MDM4 complex is characterized by a relatively high free energy, through a single energy barrier, and by a lifetime in the order of tens of seconds. New insights into the MDM2-MDM4 interaction could be highly important for developing innovative anticancer drugs focused on p53 reactivation.
Biophysics and Nanoscience Centre, Department DEB, Università della Tuscia, Viterbo, Italy.
Ilaria
I
Moscetti
Department of Endocrinology and Metabolism, Università Cattolica di Roma, Roma, Italy; Institute of Cell Biology and Neurobiology, Consiglio Nazionale delle Ricerche (CNR), Roma, Italy.
Emanuela
E
Teveroni
Institute of Cell Biology and Neurobiology, Consiglio Nazionale delle Ricerche (CNR), Roma, Italy.
Fabiola
F
Moretti
Biophysics and Nanoscience Centre, Department DEB, Università della Tuscia, Viterbo, Italy.
Anna Rita
AR
Bizzarri
Biophysics and Nanoscience Centre, Department DEB, Università della Tuscia, Viterbo, Italy.
Salvatore
S
Cannistraro
D018797:Cell Cycle Proteins; D006801:Humans; D055767:Immobilized Proteins / Q000378:metabolism; D007700:Kinetics; D018625:Microscopy, Atomic Force; D009687:Nuclear Proteins / Q000378:metabolism*; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051736:Proto-Oncogene Proteins c-mdm2 / Q000378:metabolism*; D013057:Spectrum Analysis / Q000379:methods*; D020349:Surface Plasmon Resonance / Q000379:methods*
D018797:Cell Cycle Proteins; D055767:Immobilized Proteins; C482775:MDM4 protein, human; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D051736:Proto-Oncogene Proteins c-mdm2
MDM2; MDM4; atomic force spectroscopy; surface plasmon resonance