Thermodynamics of the unfolding of the cold-shock protein from Thermotoga maritima.


Abstract

Proteins from (hyper-)thermophiles are known to exhibit high intrinsic stabilities. Commonly, their thermodynamic characterization is impeded by irreversible side reactions of the thermal analysis or calorimetrical problems. Small single-domain proteins are suitable candidates to overcome these obstacles. Here, the thermodynamics of the thermal denaturation of the recombinant cold-shock protein (Csp) from the hyperthermophilic bacterium Thermotoga maritima (Tm) was studied by differential scanning calorimetry. The unfolding transition can be described over a broad pH range (3.5-8.5) by a reversible two-state process. Maximum stability (DeltaG (25 degrees C)=6.5 kcal/mol) was observed at pH 5-6 where Tm Csp unfolds with a melting temperature at 95 degrees C. The heat capacity difference between the native and the denatured states is 1.1(+/-0.1) kcal/(mol K). At pH 7, thermal denaturation occurs at 82 degrees C. The corresponding free energy profile has its maximum at 30 degrees C with DeltaGN-->U=4.8(+/-0.5) kcal/mol. At the optimal growth temperature of T. maritima (80 degrees C), Tm Csp in the absence of ligands is only marginally stable, with a free energy of stabilization not far beyond the thermal energy. With the known stabilizing effect of nucleic acids in mind, this suggests a highly dynamical interaction of Tm Csp with its target molecules. Study holds ProTherm entries: 5478, 5479, 5480, 5481, 5482, 5483, 5484, 5485, 5486, 5487, 5488, 5489, 5490, 5491, 5492, 5493, 5494, 5495, 5496, 5497, 5498, 5499 Extra Details: cold-shock protein; differential scanning calorimetry(DSC);,hyperthermophiles; stability; Thermotoga maritima

Submission Details

ID: nqSLd7xn

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:30 p.m.

Version: 1

Publication Details
Wassenberg D;Welker C;Jaenicke R,J. Mol. Biol. (1999) Thermodynamics of the unfolding of the cold-shock protein from Thermotoga maritima. PMID:10339416
Additional Information

Study Summary

Number of data points 84
Proteins Cold shock-like protein
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: dCp pH:8.9, buffers:borate-NaOH: 20 mM ; Experimental Assay: dHcal pH:8.9, buffers:borate-NaOH: 20 mM ; Experimental Assay: Tm pH:8.9, buffers:borate-NaOH: 20 mM ; Experimental Assay: dCp pH:8.5, buffers:borate-NaOH: 20 mM ; Experimental Assay: dHcal pH:8.5, buffers:borate-NaOH: 20 mM ; Experimental Assay: Tm pH:8.5, buffers:borate-NaOH: 20 mM ; Experimental Assay: dHvH pH:8.5, buffers:borate-NaOH: 20 mM ; Experimental Assay: dCp pH:8.4, buffers:borate-NaOH: 20 mM ; Experimental Assay: dHcal pH:8.4, buffers:borate-NaOH: 20 mM ; Experimental Assay: Tm pH:8.4, buffers:borate-NaOH: 20 mM ; Experimental Assay: dCp pH:8.1, buffers:borate-NaOH: 20 mM ; Experimental Assay: dHcal pH:8.1, buffers:borate-NaOH: 20 mM ; Experimental Assay: Tm pH:8.1, buffers:borate-NaOH: 20 mM ; Experimental Assay: dHvH pH:8.1, buffers:borate-NaOH: 20 mM ; Experimental Assay: dCp pH:8.0, buffers:borate-NaOH: 20 mM ; Experimental Assay: dHcal pH:8.0, buffers:borate-NaOH: 20 mM ; Experimental Assay: Tm pH:8.0, buffers:borate-NaOH: 20 mM ; Experimental Assay: dHvH pH:8.0, buffers:borate-NaOH: 20 mM ; Experimental Assay: dCp pH:7.4, buffers:Sodium phosphate: 20 mM ; Experimental Assay: dHcal pH:7.4, buffers:Sodium phosphate: 20 mM ; Experimental Assay: Tm pH:7.4, buffers:Sodium phosphate: 20 mM ; Experimental Assay: dHvH pH:7.4, buffers:Sodium phosphate: 20 mM ; Experimental Assay: dCp pH:7.5, buffers:Sodium phosphate: 10 mM ; Experimental Assay: dHcal pH:7.5, buffers:Sodium phosphate: 10 mM ; Experimental Assay: Tm pH:7.5, buffers:Sodium phosphate: 10 mM ; Experimental Assay: dHvH pH:7.5, buffers:Sodium phosphate: 10 mM ; Experimental Assay: dCp pH:7.5, buffers:Sodium phosphate: 20 mM ; Experimental Assay: dHcal pH:7.5, buffers:Sodium phosphate: 20 mM ; Experimental Assay: Tm pH:7.5, buffers:Sodium phosphate: 20 mM ; Experimental Assay: dHvH pH:7.5, buffers:Sodium phosphate: 20 mM ; Experimental Assay: dCp pH:7.25, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dHcal pH:7.25, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: Tm pH:7.25, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dHvH pH:7.25, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dCp pH:7.2, buffers:Sodium phosphate: 20 mM ; Experimental Assay: dHcal pH:7.2, buffers:Sodium phosphate: 20 mM ; Experimental Assay: Tm pH:7.2, buffers:Sodium phosphate: 20 mM ; Experimental Assay: dHvH pH:7.2, buffers:Sodium phosphate: 20 mM ; Experimental Assay: dCp buffers:Sodium cacodylate-HCl: 20 mM, pH:7.0 ; Experimental Assay: dHcal buffers:Sodium cacodylate-HCl: 20 mM, pH:7.0 ; Experimental Assay: Tm buffers:Sodium cacodylate-HCl: 20 mM, pH:7.0 ; Experimental Assay: dHvH buffers:Sodium cacodylate-HCl: 20 mM, pH:7.0 ; Experimental Assay: dCp buffers:Sodium cacodylate-HCl: 20 mM, pH:6.9 ; Experimental Assay: dHcal buffers:Sodium cacodylate-HCl: 20 mM, pH:6.9 ; Experimental Assay: Tm buffers:Sodium cacodylate-HCl: 20 mM, pH:6.9 ; Experimental Assay: dHvH buffers:Sodium cacodylate-HCl: 20 mM, pH:6.9 ; Experimental Assay: dCp pH:6.75, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dHcal pH:6.75, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: Tm pH:6.75, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dHvH pH:6.75, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dCp buffers:Sodium cacodylate-HCl: 20 mM, pH:6.5 ; Experimental Assay: dHcal buffers:Sodium cacodylate-HCl: 20 mM, pH:6.5 ; Experimental Assay: Tm buffers:Sodium cacodylate-HCl: 20 mM, pH:6.5 ; Experimental Assay: dHvH buffers:Sodium cacodylate-HCl: 20 mM, pH:6.5 ; Experimental Assay: dCp buffers:Sodium cacodylate-HCl: 20 mM, pH:6.35 ; Experimental Assay: dHcal buffers:Sodium cacodylate-HCl: 20 mM, pH:6.35 ; Experimental Assay: Tm buffers:Sodium cacodylate-HCl: 20 mM, pH:6.35 ; Experimental Assay: dHvH buffers:Sodium cacodylate-HCl: 20 mM, pH:6.35 ; Experimental Assay: dCp buffers:Sodium cacodylate-HCl: 20 mM, pH:6.1 ; Experimental Assay: dHcal buffers:Sodium cacodylate-HCl: 20 mM, pH:6.1 ; Experimental Assay: Tm buffers:Sodium cacodylate-HCl: 20 mM, pH:6.1 ; Experimental Assay: dHvH buffers:Sodium cacodylate-HCl: 20 mM, pH:6.1 ; Experimental Assay: dCp pH:6.0, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dHcal pH:6.0, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: Tm pH:6.0, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dHvH pH:6.0, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dCp pH:5.7, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dHcal pH:5.7, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: Tm pH:5.7, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dHvH pH:5.7, buffers:Sodium cacodylate-HCl: 20 mM ; Experimental Assay: dCp pH:4.8, buffers:Sodium acetate: 20 mM ; Experimental Assay: dHcal pH:4.8, buffers:Sodium acetate: 20 mM ; Experimental Assay: Tm pH:4.8, buffers:Sodium acetate: 20 mM ; Experimental Assay: dHvH pH:4.8, buffers:Sodium acetate: 20 mM ; Experimental Assay: dCp pH:4.5, buffers:Sodium acetate: 20 mM ; Experimental Assay: dHcal pH:4.5, buffers:Sodium acetate: 20 mM ; Experimental Assay: Tm pH:4.5, buffers:Sodium acetate: 20 mM ; Experimental Assay: dCp pH:4.25, buffers:Sodium acetate: 20 mM ; Experimental Assay: dHcal pH:4.25, buffers:Sodium acetate: 20 mM ; Experimental Assay: Tm pH:4.25, buffers:Sodium acetate: 20 mM ; Experimental Assay: dHvH pH:4.25, buffers:Sodium acetate: 20 mM ; Experimental Assay: dCp pH:4.0, buffers:Sodium acetate: 20 mM ; Experimental Assay: dHcal pH:4.0, buffers:Sodium acetate: 20 mM ; Experimental Assay: Tm pH:4.0, buffers:Sodium acetate: 20 mM
Libraries Mutations for sequence MRGKVKWFDSKKGYGFITKDEGGDVFVHWSAIEMEGFKTLKEGQVVEFEIQEGKKGPQAAHVKVVE

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
1G6P 2001-11-07 SOLUTION NMR STRUCTURE OF THE COLD SHOCK PROTEIN FROM THE HYPERTHERMOPHILIC BACTERIUM THERMOTOGA MARITIMA

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Cold shock-like protein O54310 CSP_THEMA