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Wide-line NMR and DSC studies on intrinsically disordered p53 transactivation domain and its helically pre-structured segment - PubMed

Wide-line NMR and DSC studies on intrinsically disordered p53 transactivation domain and its helically pre-structured segment

Peter Tompa et al. BMB Rep. 2016 Sep.

Abstract

Wide-line 1H NMR intensity and differential scanning calorimetry measurements were carried out on the intrinsically disordered 73-residue full transactivation domain (TAD) of the p53 tumor suppressor protein and two peptides: one a wild type p53 TAD peptide with a helix pre-structuring property, and a mutant peptide with a disabled helix-forming propensity. Measurements were carried out in order to characterize their water and ion binding characteristics. By quantifying the number of hydrate water molecules, we provide a microscopic description for the interactions of water with a wild-type p53 TAD and two p53 TAD peptides. The results provide direct evidence that intrinsically disordered proteins (IDPs) and a less structured peptide not only have a higher hydration capacity than globular proteins, but are also able to bind a larger amount of charged solute ions. [BMB Reports 2016; 49(9): 497-501].

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Figures

Fig. 1.
Fig. 1.. Hydration measured by the 1H-NMR signal intensities of the mobile water for (A) 50 mg/ml p53 TAD (blue circles) and 50 mg/ml BSA (black circles) dissolved in water. The T = −4℃ data for the helices (green and red circles) are the same as in graph (B). (B) 30 mg/ml p53 TAD helix (green circles) and 30 mg/ml mutant helix (red circles) dissolved in water. The lines are guides to the eye.
Fig. 2.
Fig. 2.. Hydration measured by the 1H-NMR signal intensities of the mobile water for (A) 50 mg/ml p53 TAD (blue circles) and 50 mg/ml BSA (black circles) dissolved in Tris-buffer. The T = −4℃ data for the helix peptides (green and red circles) are the same as in graph (B). (B) 30 mg/ml wild type p53 TAD helix peptide (green circles) and 30 mg/ml mutant helix peptide (red circles) dissolved in Tris-buffer; alternately, the hydration given in units of mole mobile water per mole peptide. Tris-buffer: 150 mM NaCl, 50 mM Tris, 1 mM EDTA, pH 7.5. The lines are guides to the eye.
Fig. 3.
Fig. 3.. DSC curves measured in double-distilled water (black line), wild type helix peptide (residues 15-29 of p53 TAD) of concentrations 25 mg/cm3 (blue line) and 12.5 mg/cm3 (red line) dissolved in Tris-buffer (150 mM NaCl, 50 mM Tris, 1 mM EDTA, pH 7.5). The endotherm peak at −25℃ (red line) represents the melting of the eutectic phase formed by that fraction of the NaCl which was not bound by the peptide.
Fig. 4.
Fig. 4.. Enthalpy changes associated with the eutectic melting of the H2O-NaCl system measured as a function of protein concentration for full p53 TAD (blue squares), wild type p53 TAD helix peptide (green circles) and mutant peptide (red circle) dissolved in the buffer solution of 150 mM NaCl, 50 mM Tris, 1 mM EDTA, pH 7.5. The lines are guides to the eye.

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