A thermodynamic study of the 434-repressor N-terminal domain and of its covalently linked dimers.


Abstract

The isolated N-terminal 1-69 domain of the 434-phage repressor, R69, and its covalently linked (head-to-tail and tail-to-tail) dimers have been studied by differential scanning microcalorimetry (DSC) and CD. At neutral solvent conditions the R69 domain maintains its native structure, both in isolated form and within the dimers. The stability of the domain depends highly upon pH within the acidic range, thus at pH 2 and low ionic strength R69 is already partially unfolded at room temperature. The thermodynamic parameters of unfolding calculated from the DSC data are typical for small globular proteins. At neutral pH and moderate ionic strength, the domains of the dimers behave as two independent units with unfolding parameters similar to those of the isolated domain, which means that linking two R69 domains, either by a long peptide linker or by a designed C-terminal disulfide bridge, does not induce any cooperation between them. Study holds ProTherm entries: 5996, 5997, 5998, 5999, 6000, 6001, 6002, 6003, 6004, 6005, 6006, 6007, 6008, 6009, 6010, 6011, 6012, 6013, 6014, 6015, 6016, 6017, 6018, 6019, 6020, 6021, 6022, 6023, 6024, 6025, 6026, 6027, 6028, 6029, 6030, 6031, 6032, 6033, 6034, 6035 Extra Details: R69: isolated 1-69 N-terminal fragment of the 434-phage repressor thermal stability; differential scanning microcalorimetry;,circular dichroism; domain stability; interprotein stability; domain interactions

Submission Details

ID: fwgBGeN73

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:31 p.m.

Version: 1

Publication Details
Ruiz-Sanz J;Simoncsits A;Törö I;Pongor S;Mateo PL;Filimonov VV,Eur. J. Biochem. (1999) A thermodynamic study of the 434-repressor N-terminal domain and of its covalently linked dimers. PMID:10429210
Additional Information

Study Summary

Number of data points 70
Proteins Regulatory protein cro ; Regulatory protein cro
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: dG ionic:NaCl: 200 mM, buffers:glycine: 20 mM, pH:2.0 ; Experimental Assay: dG ionic:: , buffers:glycine: 20 mM, pH:2.0 ; Experimental Assay: dHcal ionic:NaCl: 200 mM, buffers:glycine: 20 mM, pH:2.0 ; Experimental Assay: Tm ionic:NaCl: 200 mM, buffers:glycine: 20 mM, pH:2.0 ; Experimental Assay: dHcal ionic:: , buffers:glycine: 20 mM, pH:2.0 ; Experimental Assay: Tm ionic:: , buffers:glycine: 20 mM, pH:2.0 ; Experimental Assay: dG ionic:NaCl: 200 mM, pH:3.0, buffers:glycine: 20 mM ; Experimental Assay: dG ionic:: , pH:3.0, buffers:glycine: 20 mM ; Experimental Assay: dHcal ionic:NaCl: 200 mM, pH:3.0, buffers:glycine: 20 mM ; Experimental Assay: Tm ionic:NaCl: 200 mM, pH:3.0, buffers:glycine: 20 mM ; Experimental Assay: dHcal ionic:: , pH:3.0, buffers:glycine: 20 mM ; Experimental Assay: Tm ionic:: , pH:3.0, buffers:glycine: 20 mM ; Experimental Assay: dG ionic:NaCl: 200 mM, pH:4.0, buffers:Sodium acetate: 20 mM ; Experimental Assay: dG buffers:Sodium acetate: 20 mM, pH:4.0, ionic:: ; Experimental Assay: dHcal ionic:NaCl: 200 mM, pH:4.0, buffers:Sodium acetate: 20 mM ; Experimental Assay: Tm ionic:NaCl: 200 mM, pH:4.0, buffers:Sodium acetate: 20 mM ; Experimental Assay: dHcal buffers:Sodium acetate: 20 mM, pH:4.0, ionic:: ; Experimental Assay: Tm buffers:Sodium acetate: 20 mM, pH:4.0, ionic:: ; Experimental Assay: dG ionic:NaCl: 200 mM, pH:4.5, buffers:Sodium acetate: 20 mM ; Experimental Assay: dG buffers:Sodium acetate: 20 mM, pH:4.5, ionic:: ; Experimental Assay: dHcal ionic:NaCl: 200 mM, pH:4.5, buffers:Sodium acetate: 20 mM ; Experimental Assay: Tm ionic:NaCl: 200 mM, pH:4.5, buffers:Sodium acetate: 20 mM ; Experimental Assay: dHcal buffers:Sodium acetate: 20 mM, pH:4.5, ionic:: ; Experimental Assay: Tm buffers:Sodium acetate: 20 mM, pH:4.5, ionic:: ; Experimental Assay: dG ionic:NaCl: 200 mM, buffers:PIPES: 20 mM, pH:7.0 ; Experimental Assay: dG ionic:: , buffers:PIPES: 20 mM, pH:7.0 ; Experimental Assay: dHcal ionic:NaCl: 200 mM, buffers:PIPES: 20 mM, pH:7.0 ; Experimental Assay: Tm ionic:NaCl: 200 mM, buffers:PIPES: 20 mM, pH:7.0 ; Experimental Assay: dHcal ionic:: , buffers:PIPES: 20 mM, pH:7.0 ; Experimental Assay: Tm ionic:: , buffers:PIPES: 20 mM, pH:7.0
Libraries Mutations for sequence MQTLSERLKKRRIALKMTQTELATKAGVKQQSIQLIEAGVTKRPRFLFEIAMALNCDPVWLQYGTKRGKAA

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
1ZUG 1997-07-07 STRUCTURE OF PHAGE 434 CRO PROTEIN, NMR, 20 STRUCTURES
2CRO 1989-10-15 2.35 STRUCTURE OF PHAGE 434 CRO PROTEIN AT 2.35 ANGSTROMS RESOLUTION
3CRO 1991-10-15 2.5 THE PHAGE 434 CRO/OR1 COMPLEX AT 2.5 ANGSTROMS RESOLUTION

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Regulatory protein cro P03036 RCRO_BP434