2024-03-28T19:21:41Zhttps://www.tdx.cat/oai/requestoai:www.tdx.cat:10803/28192017-09-02T07:13:52Zcom_10803_1col_10803_108
nam a 5i 4500
Complexos proteics
Dianes terapèutiques
Interaccions proteïna-proteïna
Tetramerització
Proteïna p53
Use of calix[4]arenes to recover the self-assembly ability of mutated p53 tetramerization domains
[Barcelona] :
Universitat de Barcelona,
2011
Accés lliure
http://hdl.handle.net/10803/2819
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AAMMDDs2011 sp ||||fsm||||0|| 0 eng|c
9788469157725
Gordo Villoslada, Susana,
autor
Tesi
Doctorat
Universitat de Barcelona. Departament de Química Orgànica
2008
Universitat de Barcelona. Departament de Química Orgànica
Tesis i dissertacions electròniques
Giralt Lledó, Ernest,
supervisor acadèmic
TDX
Protein-protein interactions are essential in biological processes and thus, they have become very promising therapeutic targets. Nevertheless, the artificial modulation of protein complexes remains a challenge. Since most work to date has been focused on the inhibition of protein-protein interactions, there is little precedent on the design of molecules which can induce, stabilize or recover the oligomerization state of proteins. In this context, the system comprised of the tetramerization domain of protein p53 (p53TD) and its oncogenic mutants with defective oligomerization properties is an outstanding case of study for the design and evaluation of molecules which can recover the tetrameric structure. <br/>Through a collaboration with Prof. Javier de Mendoza, a family of para-guanidinomethyl-calix[4]arenes able to interact simultaneously with the four monomers of the p53 tetramerization domain was rationally designed; hence, the interaction with these compounds would stabilize the whole tetrameric assembly. <br/>In order to evaluate experimentally said calix[4]arenes, three natural oncogenic mutants of the p53TD with defective assembly abilities were biosynthesized. Namely, they are: G334V, R337H and L344P. <br/>Once synthesized and purified the calix[4]arenes compounds, their molecular recognition properties were tested through a battery of biophysical techniques, including nuclear magnetic resonances (on both the protein and the ligand), circular dichroism, differential scanning calorimetry, crystallography, mass spectrometry and chemical cross-linking. The results clearly show that these calix[4]arenes interact with the proteins as intended and, the most important, they can thermally and kinetically stabilize the tetrameric state. <br/>These results are the perfect evidence of the proof-of-concept initially sought: a little synthetic ligand can stabilize the oligomeric state of proteins which are structurally defective. In addition, the study of several ligands with different functionalizations provides further understanding about the basis of molecular recognition events. On the one hand, the guanidinium group has a vital role for high affinity interactions. On the other hand, structural flexibility, in both the protein and the ligand, enables the molecules to adopt the optimal conformation for the tightest interaction, thereby underscoring the ambiguous and unpredictable role of the entropy in interaction processes.
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