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
1RPR 1994-01-31 THE STRUCTURE OF COLE1 ROP IN SOLUTION
1NKD 1999-03-23 1.09 ATOMIC RESOLUTION (1.07 ANGSTROMS) STRUCTURE OF THE ROP MUTANT <2AA>
4DO2 2013-02-13 1.4 Crystal Structure of the Rop protein mutant D30P/A31G at resolution 1.4 resolution.
1RPO 1995-02-14 1.4 RESTORED HEPTAD PATTERN CONTINUITY DOES NOT ALTER THE FOLDING OF A 4-ALPHA-HELICAL BUNDLE
2IJK 2007-10-16 1.55 Structure of a Rom protein dimer at 1.55 angstrom resolution
1ROP 1992-07-15 1.7 STRUCTURE OF THE COL*E1 ROP PROTEIN AT 1.7 ANGSTROMS RESOLUTION
2IJH 2007-10-16 1.8 Crystal structure analysis of ColE1 ROM mutant F14W
1B6Q 1999-07-09 1.8 ALANINE 31 PROLINE MUTANT OF ROP PROTEIN
1GTO 1997-01-27 1.82 HIGH RESOLUTION STRUCTURE OF A HYPERSTABLE HELICAL BUNDLE PROTEIN MUTANT
2IJJ 2007-10-16 1.9 Crystal structure analysis of ColE1 ROM mutant F14Y
1GMG 2002-09-12 1.9 ALANINE 31 PROLINE MUTANT OF ROP PROTEIN, MONOCLINIC FORM
1F4N 2001-01-10 1.9 C2 CRYSTAL STRUCTURE OF ALA2ILE2-6, A VERSION OF ROP WITH A REPACKED HYDROPHOBIC CORE AND A NEW FOLD.
3K79 2010-02-02 1.96 C38A, C52V Cysteine-Free Variant of Rop (Rom)
1QX8 2004-09-28 2.02 Crystal structure of a five-residue deletion mutant of the Rop protein
1F4M 2001-01-10 2.25 P3(2) CRYSTAL STRUCTURE OF ALA2ILE2-6, A VERSION OF ROP WITH A REPACKED HYDROPHOBIC CORE AND A NEW FOLD.
2IJI 2007-10-16 2.3 Structure of F14H mutant of ColE1 Rom protein
2GHY 2006-05-30 2.5 Novel Crystal Form of the ColE1 Rom Protein
1YO7 2005-02-15 2.8 Re-engineering topology of the homodimeric ROP protein into a single-chain 4-helix bundle

Relevant UniProtKB Entries

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