Slow unfolding and refolding kinetics of the mesophilic Rop wild-type protein in the transition range.


Abstract

We describe the guanidinium hydrochloride induced folding kinetics of the four-helix-bundle protein Rop wild-type (wt) under equilibrium conditions at three temperatures. The choice of appropriate denaturant conditions inside the transition range permitted, in combination with equilibrium transition curves, the determination of both unfolding and refolding rate constants. The ratio of the rate constants at zero denaturant concentration provided equilibrium constants and standard free energy changes that are in good agreement with values obtained in previous differential scanning calorimetry studies. The DeltaG0D values for 19, 25 and 40 degrees C calculated from the present kinetic studies are, respectively, 66.8, 70.8 and 57.2 kJ.mol-1. The unfolding reactions are extremely slow under these conditions. Equilibrium was reached only after 18, 12 and 6 days at 19, 25 and 40 degrees C. These results demonstrate that for Rop wt high stability correlates with slow folding kinetics. Study holds ProTherm entries: 15091, 15092, 15093 Extra Details: Additive:EDTA 1mM Rop protein; folding kinetics; slow equilibrium folding; stability

Submission Details

ID: 9sV5aQoG

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:45 p.m.

Version: 1

Publication Details
Rosengarth A;Rösgen J;Hinz HJ,Eur. J. Biochem. (1999) Slow unfolding and refolding kinetics of the mesophilic Rop wild-type protein in the transition range. PMID:10491149
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
1B6Q 1999-01-16T00:00:00+0000 1.8 ALANINE 31 PROLINE MUTANT OF ROP PROTEIN
1F4M 2000-06-08T00:00:00+0000 2.25 P3(2) CRYSTAL STRUCTURE OF ALA2ILE2-6, A VERSION OF ROP WITH A REPACKED HYDROPHOBIC CORE AND A NEW FOLD.
1F4N 2000-06-08T00:00:00+0000 1.9 C2 CRYSTAL STRUCTURE OF ALA2ILE2-6, A VERSION OF ROP WITH A REPACKED HYDROPHOBIC CORE AND A NEW FOLD.
1GMG 2001-09-13T00:00:00+0000 1.9 ALANINE 31 PROLINE MUTANT OF ROP PROTEIN, MONOCLINIC FORM
1GTO 1996-04-23T00:00:00+0000 1.82 HIGH RESOLUTION STRUCTURE OF A HYPERSTABLE HELICAL BUNDLE PROTEIN MUTANT
1NKD 1997-09-23T00:00:00+0000 1.09 ATOMIC RESOLUTION (1.07 ANGSTROMS) STRUCTURE OF THE ROP MUTANT <2AA>
1QX8 2003-09-04T00:00:00+0000 2.02 Crystal structure of a five-residue deletion mutant of the Rop protein
1ROP 1991-04-02T00:00:00+0000 1.7 STRUCTURE OF THE COL*E1 ROP PROTEIN AT 1.7 ANGSTROMS RESOLUTION
1RPO 1994-08-25T00:00:00+0000 1.4 RESTORED HEPTAD PATTERN CONTINUITY DOES NOT ALTER THE FOLDING OF A 4-ALPHA-HELICAL BUNDLE
1RPR 1991-10-09T00:00:00+0000 0 THE STRUCTURE OF COLE1 ROP IN SOLUTION

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Regulatory protein rop P03051 ROP_ECOLX