DOI: 10.1073/pnas.94.26.14338. Thermodynamic stability of wild-type and mutant p53 core domain

Thermodynamic stability of wild-type and mutant p53 core domain

10.1073/pnas.94.26.14338

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Some 50% of human cancers are associated with mutations in the core domain of the tumor suppressor p53. Many mutations are thought just to destabilize the protein. To assess this and the possibility of rescue, we have set up a system to analyze the stability of the core domain and its mutants. The use of differential scanning calorimetry or spectroscopy to measure its melting temperature leads to irreversible denaturation and aggregation and so is useful as only a qualitative guide to stability. There are excellent two-state denaturation curves on the addition of urea that may be analyzed quantitatively. One Zn 2+ ion remains tightly bound in the holo-form of p53 throughout the denaturation curve. The stability of wild type is 6.0 kcal (1 kcal = 4.18 kJ)/mol at 25°C and 9.8 kcal/mol at 10°C. The oncogenic mutants R175H, C242S, R248Q, R249S, and R273H are destabilized by 3.0, 2.9, 1.9, 1.9, and 0.4 kcal/mol, respectively. Under certain denaturing conditions, the wild-type domain forms an aggregate that is relatively highly fluorescent at 340 nm on excitation at 280 nm. The destabilized mutants give this fluorescence under milder denaturation conditions.

Bibliography

Bullock, A. N., Henckel, J., DeDecker, B. S., Johnson, C. M., Nikolova, P. V., Proctor, M. R., Lane, D. P., & Fersht, A. R. (1997). Thermodynamic stability of wild-type and mutant p53 coreâdomain. Proceedings of the National Academy of Sciences, 94(26), 14338â14342.

Authors 8

Alex N. Bullock (first)
Julia Henckel (additional)
Brian S. DeDecker (additional)
Christopher M. Johnson (additional)
Penka V. Nikolova (additional)
Mark R. Proctor (additional)
David P. Lane (additional)
Alan R. Fersht (additional)

References 22 Referenced 362

10.1038/358015a0
M B Kastan, O Onyekwere, D Sidransky, B Vogelstein, R W Craig Cancer Res 51, 6304–6311 (1991). / Cancer Res by Kastan M B (1991)
10.1038/38525
10.1016/S0021-9258(17)36921-1
10.1038/387296a0
10.1101/gad.7.12b.2556
10.1126/science.8023157
N Rolley, S Butcher, J Milner Oncogene 11, 763–770 (1995). / Oncogene by Rolley N (1995)
10.1074/jbc.271.41.25468
J Milner, A C Cook, M Sheldon Oncogene 1, 453–455 (1987). / Oncogene by Milner J (1987)
10.1021/bi00067a022
10.1016/0022-2836(92)90562-X
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10.1016/0003-2697(85)90409-9
10.1016/0959-440X(93)90205-Y
10.1107/S0021889891004399

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:31 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 2:24 p.m.)
Indexed	1 month ago (July 30, 2025, 10:26 a.m.)
Issued	27 years, 8 months ago (Dec. 23, 1997)
Published	27 years, 8 months ago (Dec. 23, 1997)
Published Online	27 years, 8 months ago (Dec. 23, 1997)
Published Print	27 years, 8 months ago (Dec. 23, 1997)

Funders 0

None

BibTeX

@article{Bullock_1997, title={Thermodynamic stability of wild-type and mutant p53 core domain}, volume={94}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.94.26.14338}, DOI={10.1073/pnas.94.26.14338}, number={26}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Bullock, Alex N. and Henckel, Julia and DeDecker, Brian S. and Johnson, Christopher M. and Nikolova, Penka V. and Proctor, Mark R. and Lane, David P. and Fersht, Alan R.}, year={1997}, month=dec, pages={14338–14342} }

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