000129663 001__ 129663 000129663 005__ 20241125101135.0 000129663 0247_ $$2doi$$a10.3390/molecules28227578 000129663 0248_ $$2sideral$$a135859 000129663 037__ $$aART-2023-135859 000129663 041__ $$aeng 000129663 100__ $$aRizzuti, Bruno 000129663 245__ $$aConformational Stability of the N-Terminal Region of MDM2 000129663 260__ $$c2023 000129663 5060_ $$aAccess copy available to the general public$$fUnrestricted 000129663 5203_ $$aMDM2 is an E3 ubiquitin ligase which is crucial for the degradation and inhibition of the key tumor-suppressor protein p53. In this work, we explored the stability and the conformational features of the N-terminal region of MDM2 (N-MDM2), through which it binds to the p53 protein as well as other protein partners. The isolated domain possessed a native-like conformational stability in a narrow pH range (7.0 to 10.0), as shown by intrinsic and 8-anilinonapthalene-1-sulfonic acid (ANS) fluorescence, far-UV circular dichroism (CD), and size exclusion chromatography (SEC). Guanidinium chloride (GdmCl) denaturation followed by intrinsic and ANS fluorescence, far-UV CD and SEC at physiological pH, and differential scanning calorimetry (DSC) and thermo-fluorescence experiments showed that (i) the conformational stability of isolated N-MDM2 was very low; and (ii) unfolding occurred through the presence of several intermediates. The presence of a hierarchy in the unfolding intermediates was also evidenced through DSC and by simulating the unfolding process with the help of computational techniques based on constraint network analysis (CNA). We propose that the low stability of this protein is related to its inherent flexibility and its ability to interact with several molecular partners through different routes. 000129663 536__ $$9info:eu-repo/grantAgreement/ES/DGA/B25-20R$$9info:eu-repo/grantAgreement/ES/DGA/E45-20R$$9info:eu-repo/grantAgreement/ES/ISCIII/PI18-0394$$9info:eu-repo/grantAgreement/ES/ISCIII/PI21-00394$$9info:eu-repo/grantAgreement/ES/MICINN/AEI/PID2021-127296OB-I00 000129663 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/ 000129663 590__ $$a4.2$$b2023 000129663 592__ $$a0.744$$b2023 000129663 591__ $$aCHEMISTRY, MULTIDISCIPLINARY$$b77 / 231 = 0.333$$c2023$$dQ2$$eT2 000129663 593__ $$aAnalytical Chemistry$$c2023$$dQ1 000129663 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b88 / 313 = 0.281$$c2023$$dQ2$$eT1 000129663 593__ $$aPharmaceutical Science$$c2023$$dQ1 000129663 593__ $$aChemistry (miscellaneous)$$c2023$$dQ1 000129663 593__ $$aOrganic Chemistry$$c2023$$dQ2 000129663 593__ $$aPhysical and Theoretical Chemistry$$c2023$$dQ2 000129663 593__ $$aDrug Discovery$$c2023$$dQ2 000129663 593__ $$aMedicine (miscellaneous)$$c2023$$dQ2 000129663 593__ $$aMolecular Medicine$$c2023$$dQ3 000129663 594__ $$a7.4$$b2023 000129663 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion 000129663 700__ $$0(orcid)0000-0001-5664-1729$$aAbian, Olga$$uUniversidad de Zaragoza 000129663 700__ $$0(orcid)0000-0001-5702-4538$$aVelazquez-Campoy, Adrián$$uUniversidad de Zaragoza 000129663 700__ $$aNeira, José L. 000129663 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole. 000129663 773__ $$g28, 22 (2023), 7578 [21 pp.]$$pMolecules (Basel, Online)$$tMolecules$$x1420-3049 000129663 8564_ $$s3805555$$uhttps://zaguan.unizar.es/record/129663/files/texto_completo.pdf$$yVersión publicada 000129663 8564_ $$s2695482$$uhttps://zaguan.unizar.es/record/129663/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada 000129663 909CO $$ooai:zaguan.unizar.es:129663$$particulos$$pdriver 000129663 951__ $$a2024-11-22-12:00:24 000129663 980__ $$aARTICLE