Thermodynamic analysis of the structural stability of the tetrameric oligomerization domain of p53 tumor suppressor.


Abstract

The structural stability of an amino acid fragment containing the oligomerization domain (residues 303-366) of the tumor suppressor p53 has been studied using high-precision differential scanning calorimetry (DSC) and circular dichroism spectroscopy (CD). Previous NMR solution structural determinations have revealed that the fragment forms a symmetric 29.8 kDa tetramer composed of a dimer of dimers (p53tet) [Lee, W., Harvey, T. S., Yin, Y., Yau, P., Litchfield, D., & Arrowsmith, C. H. (1994) Nature Struct. Biol. 1, 877-890]. Thermal unfolding of the tetramer is reversible and can be described as a two-state transition in which the folded tetramer is converted directly to unfolded monomers (N4<==>4U). According to the DSC and CD data, the population of intermediate species consisting of folded monomers or dimers is insignificant, indicating that isolated dimeric or monomeric structures have a much lower stability than the dimer and do not become populated during thermal denaturation under the conditions studied. The transition temperature of unfolding is found to be highly dependent on protein concentration and to follow the expected behavior for a tetramer that dissociates upon unfolding. Experiments conducted at pH 4.0 in 25 mM sodium acetate at a tetramer concentration of 145.8 microM have a transition temperature (Tm) of 75.3 degrees C while at 0.5 microM the value drops to 39.2 degrees C. The enthalpy change of unfolding at 60 degrees C is 26 kcal (mol of monomer)-1 with a heat capacity change of 387 cal (K.mol of monomer)-1. The stability of p53tet is dependent on pH and salt concentration.(ABSTRACT TRUNCATED AT 250 WORDS) Study holds ProTherm entries: 5065, 5066, 5067, 5068 Extra Details:

Submission Details

ID: i5TAFMv6

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:28 p.m.

Version: 1

Publication Details
Johnson CR;Morin PE;Arrowsmith CH;Freire E,Biochemistry (1995) Thermodynamic analysis of the structural stability of the tetrameric oligomerization domain of p53 tumor suppressor. PMID:7727392
Additional Information

Study Summary

Number of data points 12
Proteins Cellular tumor antigen p53 ; Cellular tumor antigen p53
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: dCp buffers:phosphate: 25 mM, pH:7.0 ; Experimental Assay: dHcal buffers:phosphate: 25 mM, pH:7.0 ; Experimental Assay: Tm buffers:phosphate: 25 mM, pH:7.0 ; Experimental Assay: dCp pH:6.0, buffers:phosphate: 25 mM ; Experimental Assay: dHcal pH:6.0, buffers:phosphate: 25 mM ; Experimental Assay: Tm pH:6.0, buffers:phosphate: 25 mM ; Experimental Assay: dCp pH:4.0, buffers:acetate: 25 mM ; Experimental Assay: dHcal pH:4.0, buffers:acetate: 25 mM ; Experimental Assay: Tm pH:4.0, buffers:acetate: 25 mM ; Experimental Assay: dCp pH:3.0, buffers:glycine: 25 mM ; Experimental Assay: dHcal pH:3.0, buffers:glycine: 25 mM ; Experimental Assay: Tm pH:3.0, buffers:glycine: 25 mM
Libraries Mutations for sequence A:SSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNT/B:SSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNT/C:SSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNT/E:TTTCCTAGACTTGCCCAATTA/F:ATAATTGGGCAAGTCTAGGAA

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