Markov state models and NMR uncover an overlooked allosteric loop in p53 |
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Authors: | Emilia P Barros
zlem Demir Jenaro Soto Melanie J Cocco Rommie E Amaro |
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Institution: | Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA 92093 USA, Fax: +1-858-534-9645, +1-858-534-9629 ; Department of Pharmaceutical Sciences, University of California Irvine, Irvine CA 92697 USA ; Department of Molecular Biology and Biochemistry, University of California Irvine, Irvine 92697 CA USA |
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Abstract: | The tumor suppressor p53 is the most frequently mutated gene in human cancer, and thus reactivation of mutated p53 is a promising avenue for cancer therapy. Analysis of wildtype p53 and the Y220C cancer mutant long-timescale molecular dynamics simulations with Markov state models and validation by NMR relaxation studies has uncovered the involvement of loop L6 in the slowest motions of the protein. Due to its distant location from the DNA-binding surface, the conformational dynamics of this loop has so far remained largely unexplored. We observe mutation-induced stabilization of alternate L6 conformations, distinct from all experimentally-determined structures, in which the loop is both extended and located further away from the DNA-interacting surface. Additionally, the effect of the L6-adjacent Y220C mutation on the conformational landscape of the functionally-important loop L1 suggests an allosteric role to this dynamic loop and the inactivation mechanism of the mutation. Finally, the simulations reveal a novel Y220C cryptic pocket that can be targeted for p53 rescue efforts. Our approach exemplifies the power of the MSM methodology for uncovering intrinsic dynamic and kinetic differences among distinct protein ensembles, such as for the investigation of mutation effects on protein function.Wildtype and Y220C L1 and L6 loops conformational landscape, with MSM-identified L6 states highlighted on the right. |
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