Thermodynamic stability of wild-type and mutant p53 core domain.


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 Zn2+ 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 degrees C and 9.8 kcal/mol at 10 degrees 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. Study holds ProTherm entries: 3045, 3046, 3047, 3048, 3049, 3050, 3051 Extra Details: thermodynamic stability; p53 core domain; tumor suppressor;,fluorescence; calorimetry

Submission Details

ID: BAAcHpTe3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:20 p.m.

Version: 1

Publication Details
Bullock AN;Henckel J;DeDecker BS;Johnson CM;Nikolova PV;Proctor MR;Lane DP;Fersht AR,Proc. Natl. Acad. Sci. U.S.A. (1997) Thermodynamic stability of wild-type and mutant p53 core domain. PMID:9405613
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1A1U 1997-12-16T00:00:00+0000 0 SOLUTION STRUCTURE DETERMINATION OF A P53 MUTANT DIMERIZATION DOMAIN, NMR, MINIMIZED AVERAGE STRUCTURE
1AIE 1997-04-17T00:00:00+0000 1.5 P53 TETRAMERIZATION DOMAIN CRYSTAL STRUCTURE
1C26 1999-07-22T00:00:00+0000 1.7 CRYSTAL STRUCTURE OF P53 TETRAMERIZATION DOMAIN
1DT7 2000-01-11T00:00:00+0000 0 SOLUTION STRUCTURE OF THE C-TERMINAL NEGATIVE REGULATORY DOMAIN OF P53 IN A COMPLEX WITH CA2+-BOUND S100B(BB)
1GZH 2002-05-22T00:00:00+0000 2.6 Crystal structure of the BRCT domains of human 53BP1 bound to the p53 tumor supressor
1GZH 2002-05-22T00:00:00+0000 2.6 Crystal structure of the BRCT domains of human 53BP1 bound to the p53 tumor supressor
1H26 2002-07-31T00:00:00+0000 2.24 CDK2/CyclinA in complex with an 11-residue recruitment peptide from p53
1HS5 2000-12-22T00:00:00+0000 0 NMR SOLUTION STRUCTURE OF DESIGNED P53 DIMER
1JSP 2001-08-17T00:00:00+0000 0 NMR Structure of CBP Bromodomain in complex with p53 peptide
1KZY 2002-02-08T00:00:00+0000 2.5 Crystal Structure of the 53bp1 BRCT Region Complexed to Tumor Suppressor P53

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
277.5 A,B,C Cellular tumor antigen p53 Q29480 P53_EQUAS
277.79999999999995 A,B,C Cellular tumor antigen p53 P79892 P53_HORSE
272.4 A,B,C Cellular tumor antigen p53 Q9TTA1 P53_TUPBE
277.79999999999995 A,B,C Cellular tumor antigen p53 O36006 P53_MARMO
276.6 A,B,C Cellular tumor antigen p53 Q64662 P53_OTOBE
283.5 A,B,C Cellular tumor antigen p53 Q95330 P53_RABIT
290.4 A,B,C Cellular tumor antigen p53 P56424 P53_MACMU
290.4 A,B,C Cellular tumor antigen p53 P61260 P53_MACFU
290.4 A,B,C Cellular tumor antigen p53 P56423 P53_MACFA
291.6 A,B,C Cellular tumor antigen p53 P13481 P53_CHLAE
300.0 A,B,C Cellular tumor antigen p53 P04637 P53_HUMAN