Breakdown in the relationship between thermal and thermodynamic stability in an interleukin-1 beta point mutant modified in a surface loop.


Abstract

Sequence variants of the beta-barrel protein interleukin-1 beta have been analyzed for their stabilities toward irreversible thermal inactivation by monitoring the generation of light scattering aggregates on heating. The derived temperatures for the onset of aggregation (T(agg) values) correlate well with the free energies of unfolding of these proteins with the exception of one variant, Lys97-->Val (K97V), which undergoes aggregation at a temperature 7 degrees C lower than expected based on its thermodynamic stability. This lower than expected thermal stability may be due to generation of an aggregation-prone unfolding intermediate at a temperature lower than the Tm of the global transition. This hypothesis is supported by the location of residue 97 in the long 86-99 loop which has structural features suggesting it may comprise a small, independent folding unit or microdomain. The excellent correlation of thermal and thermodynamic stabilities of seven of the eight variants tested is consistent with accepted models for thermal inactivation of proteins. At the same time the poor fit of the K97V variant underscores the risk in using thermal stability data in quantitative analysis of mutational studies of the folding stability of proteins.

Submission Details

ID: T35CrH3W4

Submitter: Shu-Ching Ou

Submission Date: Oct. 23, 2018, 2:27 p.m.

Version: 1

Publication Details
Chrunyk BA;Wetzel R,Protein Eng (1993) Breakdown in the relationship between thermal and thermodynamic stability in an interleukin-1 beta point mutant modified in a surface loop. PMID:8248096
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
2KH2 2009-09-08 Solution structure of a scFv-IL-1B complex
7I1B 1992-10-15 HIGH-RESOLUTION THREE-DIMENSIONAL STRUCTURE OF INTERLEUKIN-1 BETA IN SOLUTION BY THREE-AND FOUR-DIMENSIONAL NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY
6I1B 1992-10-15 HIGH-RESOLUTION THREE-DIMENSIONAL STRUCTURE OF INTERLEUKIN-1 BETA IN SOLUTION BY THREE-AND FOUR-DIMENSIONAL NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY
4GAI 2013-02-20 1.49 Crystal structure of EBI-005, a chimera of human IL-1beta and IL-1Ra
2NVH 2006-12-12 1.53 Determination of Solvent Content in Cavities in Interleukin-1 Using Experimentally-Phased Electron Density
1L2H 2003-02-04 1.54 Crystal structure of Interleukin 1-beta F42W/W120F mutant
3POK 2011-01-19 1.7 Interleukin-1-beta LBT L3 Mutant
4G6M 2012-12-19 1.81 Crystal structure of human IL-1beta in complex with therapeutic antibody binding fragment of gevokizumab
2I1B 1990-04-15 2.0 CRYSTALLOGRAPHIC REFINEMENT OF INTERLEUKIN-1 BETA AT 2.0 ANGSTROMS RESOLUTION
4I1B 1990-04-15 2.0 FUNCTIONAL IMPLICATIONS OF INTERLEUKIN-1BETA BASED ON THE THREE-DIMENSIONAL STRUCTURE
1IOB 1996-08-17 2.0 INTERLEUKIN-1 BETA FROM JOINT X-RAY AND NMR REFINEMENT
1I1B 1990-01-15 2.0 CRYSTAL STRUCTURE OF RECOMBINANT HUMAN INTERLEUKIN-1BETA AT 2.0 ANGSTROMS RESOLUTION
4G6J 2012-12-19 2.03 Crystal structure of human IL-1beta in complex with the therapeutic antibody binding fragment of canakinumab
5I1B 1992-01-15 2.1 A COMPARISON OF THE HIGH RESOLUTION STRUCTURES OF HUMAN AND MURINE INTERLEUKIN-1B
1T4Q 2004-12-07 2.1 Interleukin 1 beta F101W
3LTQ 2011-02-16 2.1 Structure of Interleukin 1B solved by SAD using an inserted Lanthanide Binding Tag
1TOO 2004-12-07 2.1 Interleukin 1B Mutant F146W
1TWE 2004-12-07 2.1 INTERLEUKIN 1 BETA MUTANT F101Y
5MVZ 2017-02-15 2.15 Fab 4AB007 bound to Interleukin-1-beta
4GAF 2013-02-20 2.15 Crystal structure of EBI-005, a chimera of human IL-1beta and IL-1Ra, bound to human Interleukin-1 receptor type 1
5BVP 2015-09-02 2.2 The molecular mode of action and species specificity of canakinumab, a human monoclonal antibody neutralizing IL-1beta
1TP0 2005-02-08 2.2 Triple mutation in interleukin 1 beta cavity:replacement of phenylalanines with tryptophan.
1TWM 2004-12-07 2.26 Interleukin-1 Beta Mutant F146Y
9ILB 1999-01-06 2.28 HUMAN INTERLEUKIN-1 BETA
1S0L 2004-03-30 2.34 Interleukin 1 beta mutant F42W
1HIB 1994-01-31 2.4 THE STRUCTURE OF AN INTERLEUKIN-1 BETA MUTANT WITH REDUCED BIOACTIVITY SHOWS MULTIPLE SUBTLE CHANGES IN CONFORMATION THAT AFFECT PROTEIN-PROTEIN RECOGNITION
1ITB 1998-02-04 2.5 TYPE-1 INTERLEUKIN-1 RECEPTOR COMPLEXED WITH INTERLEUKIN-1 BETA
4DEP 2012-03-21 3.1 Structure of the IL-1b signaling complex
3O4O 2010-09-01 3.3 Crystal structure of an Interleukin-1 receptor complex

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
94.8 Interleukin-1 beta P46648 IL1B_CERAT
95.4 Interleukin-1 beta P79182 IL1B_MACFA
95.4 Interleukin-1 beta P51493 IL1B_MACNE
96.1 Interleukin-1 beta P48090 IL1B_MACMU
100.0 Interleukin-1 beta P01584 IL1B_HUMAN