Methionine oxidation of monomeric lambda repressor: the denatured state ensemble under nondenaturing conditions.


Abstract

Although poorly understood, the properties of the denatured state ensemble are critical to the thermodynamics and the kinetics of protein folding. The most relevant conformations to cellular protein folding are the ones populated under physiological conditions. To avoid the problem of low expression that is seen with unstable variants, we used methionine oxidation to destabilize monomeric lambda repressor and predominantly populate the denatured state under nondenaturing buffer conditions. The denatured ensemble populated under these conditions comprises conformations that are compact. Analytical ultracentrifugation sedimentation velocity experiments indicate a small increase in Stokes radius over that of the native state. A significant degree of alpha-helical structure in these conformations is detected by far-UV circular dichroism, and some tertiary interactions are suggested by near-UV circular dichroism. The characteristics of the denatured state populated by methionine oxidation in nondenaturing buffer are very different from those found in chemical denaturant. Study holds ProTherm entries: 20568 Extra Details: repressor proteins; denatured state ensemble; methionine oxidation; hydrodynamic radius; heteronuclear NMR; protein structure/folding; NMR spectroscopy; new methods; circular dichroism

Submission Details

ID: bTmfnWVC3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:52 p.m.

Version: 1

Publication Details
Chugha P;Sage HJ;Oas TG,Protein Sci. (2006) Methionine oxidation of monomeric lambda repressor: the denatured state ensemble under nondenaturing conditions. PMID:16452618
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
3KZ3 2010-02-23 1.64 A structure of a lambda repressor fragment mutant
5ZCA 2018-08-15 1.8 Crystal structure of lambda repressor (1-20) fused with maltose-binding protein
1LMB 1991-11-05 1.8 REFINED 1.8 ANGSTROM CRYSTAL STRUCTURE OF THE LAMBDA REPRESSOR-OPERATOR COMPLEX
1F39 2000-07-26 1.9 CRYSTAL STRUCTURE OF THE LAMBDA REPRESSOR C-TERMINAL DOMAIN
3WOA 2015-04-29 2.0 Crystal structure of lambda repressor (1-45) fused with maltose-binding protein
1LLI 1994-08-31 2.1 THE CRYSTAL STRUCTURE OF A MUTANT PROTEIN WITH ALTERED BUT IMPROVED HYDROPHOBIC CORE PACKING
1RIO 2004-01-27 2.3 Structure of bacteriophage lambda cI-NTD in complex with sigma-region4 of Thermus aquaticus bound to DNA
1KCA 2001-12-21 2.91 Crystal Structure of the lambda Repressor C-terminal Domain Octamer
1LRP 1989-01-09 3.2 COMPARISON OF THE STRUCTURES OF CRO AND LAMBDA REPRESSOR PROTEINS FROM BACTERIOPHAGE LAMBDA
3BDN 2008-04-15 3.91 Crystal Structure of the Lambda Repressor

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Repressor protein cI P03034 RPC1_LAMBD