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Title: Kinetic and thermodynamic effects of phosphorylation on p53 binding to MDM2
Authors: Yadahalli, Shilpa
Neira, José L.
Johnson, Christopher M.
Tan, Yaw Sing
Rowling, Pamela J. E.
Chattopadhyay, Anasuya
Itzhaki, Laura S.
Verma, Chandra Shekhar
Keywords: Phosphorylation
DRNTU::Science::Biological sciences
Thermodynamic Effects
Issue Date: 2019
Source: Yadahalli, S., Neira, J. L., Johnson, C. M., Tan, Y. S., Rowling, P. J. E., Chattopadhyay, A., . . . Itzhaki, L. S. (2019). Kinetic and thermodynamic effects of phosphorylation on p53 binding to MDM2. Scientific Reports, 9, 693-. doi:10.1038/s41598-018-36589-5
Series/Report no.: Scientific Reports
Abstract: p53 is frequently mutated in human cancers. Its levels are tightly regulated by the E3 ubiquitin ligase MDM2. The complex between MDM2 and p53 is largely formed by the interaction between the N-terminal domain of MDM2 and the N-terminal transactivation (TA) domain of p53 (residues 15–29). We investigated the kinetic and thermodynamic basis of the MDM2/p53 interaction by using wild-type and mutant variants of the TA domain. We focus on the effects of phosphorylation at positions Thr18 and Ser20 including their substitution with phosphomimetics. Conformational propensities of the isolated peptides were investigated using in silico methods and experimentally by circular dichroism and 1H-NMR in aqueous solution. Both experimental and computational analyses indicate that the p53 peptides are mainly disordered in aqueous solution, with evidence of nascent helix around the Ser20-Leu25 region. Both phosphorylation and the phosphomimetics at Thr18 result in a decrease in the binding affinity by ten- to twenty-fold when compared to the wild-type. Phosphorylation and phosphomimetics at Ser20 result in a smaller decrease in the affinity. Mutation of Lys24 and Leu25 also disrupts the interaction. Our results may be useful for further development of peptide-based drugs targeting the MDM2/p53 interaction.
DOI: 10.1038/s41598-018-36589-5
Rights: © 2019 The Author(s) (Nature Publishing Group). Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit
Fulltext Permission: open
Fulltext Availability: With Fulltext
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