Conformational Stability of the N-Terminal Region of MDM2

Rizzuti, Bruno ; Abian, Olga (Universidad de Zaragoza) ; Velazquez-Campoy, Adrián (Universidad de Zaragoza) ; Neira, José L.

Resumen: MDM2 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.

Idioma: Inglés

DOI: 10.3390/molecules28227578

Año: 2023

Publicado en: Molecules 28, 22 (2023), 7578 [21 pp.]
ISSN: 1420-3049

Factor impacto JCR: 4.2 (2023)

Categ. JCR: CHEMISTRY, MULTIDISCIPLINARY rank: 77 / 231 = 0.333 (2023) - Q2 - T2
Categ. JCR: BIOCHEMISTRY & MOLECULAR BIOLOGY rank: 88 / 313 = 0.281 (2023) - Q2 - T1

Factor impacto CITESCORE: 7.4 - Pharmaceutical Science (Q1) - Analytical Chemistry (Q1) - Physical and Theoretical Chemistry (Q1) - Organic Chemistry (Q1) - Chemistry (miscellaneous) (Q1) - Drug Discovery (Q2) - Molecular Medicine (Q2)

Factor impacto SCIMAGO: 0.744 - Analytical Chemistry (Q1) - Pharmaceutical Science (Q1) - Chemistry (miscellaneous) (Q1) - Organic Chemistry (Q2) - Physical and Theoretical Chemistry (Q2) - Drug Discovery (Q2) - Medicine (miscellaneous) (Q2) - Molecular Medicine (Q3)

Financiación: info:eu-repo/grantAgreement/ES/DGA/B25-20R
Financiación: info:eu-repo/grantAgreement/ES/DGA/E45-20R
Financiación: info:eu-repo/grantAgreement/ES/ISCIII/PI18-0394
Financiación: info:eu-repo/grantAgreement/ES/ISCIII/PI21-00394
Financiación: info:eu-repo/grantAgreement/ES/MICINN/AEI/PID2021-127296OB-I00

Tipo y forma: Artículo (Versión definitiva)

Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)