The quantitative comparison of the effects of denaturants on the stability of proteins.


Abstract

A novel quantitative comparison of denaturants involving the complete reversible unfolding of proteins is presented. Ribonuclease A was denatured with guanidinium chloride in the presence of low fixed concentrations of various partial denaturants, with the unfolding process being monitored by circular dichroism and difference spectroscopy. The major advantage of this method is that it allows a direct quantitative comparison of the effects of denaturants on the stability of proteins. The effect on the stability of ribonuclease A was shown to be linearly dependent upon the concentration of denaturant. An investigation of the constitutive ions of salts revealed that their effects were additive only in the case of salts that have no specific binding capability. This method can also be useful in detecting the specific binding of salts. Study holds ProTherm entries: 12278, 12279, 12280, 12281, 12282, 12283, 12284, 12285, 12286, 12287, 12288, 12289, 12290, 12291, 12292, 12293, 12294, 12295, 12296, 12297, 12298, 12299 Extra Details: ribonuclease A; denaturants; ions; binding

Submission Details

ID: iMttQk9G

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:43 p.m.

Version: 1

Publication Details
Thomson JA;Bigelow CC,Biochem. Cell Biol. (1986) The quantitative comparison of the effects of denaturants on the stability of proteins. PMID:3801184
Additional Information

Study Summary

Number of data points 66
Proteins Ribonuclease pancreatic ; Ribonuclease pancreatic
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: Cm ionic:LiSCN: 0.69 M ; Experimental Assay: m ionic:LiSCN: 0.69 M ; Experimental Assay: dG_H2O ionic:LiSCN: 0.69 M ; Experimental Assay: Cm ionic:LiSCN: 0.34 M ; Experimental Assay: m ionic:LiSCN: 0.34 M ; Experimental Assay: dG_H2O ionic:LiSCN: 0.34 M ; Experimental Assay: Cm ionic:LiSCN: 0.09 M ; Experimental Assay: m ionic:LiSCN: 0.09 M ; Experimental Assay: dG_H2O ionic:LiSCN: 0.09 M ; Experimental Assay: Cm ionic:NaSCN: 0.49 M ; Experimental Assay: m ionic:NaSCN: 0.49 M ; Experimental Assay: dG_H2O ionic:NaSCN: 0.49 M ; Experimental Assay: Cm ionic:NaSCN: 0.25 M ; Experimental Assay: m ionic:NaSCN: 0.25 M ; Experimental Assay: dG_H2O ionic:NaSCN: 0.25 M ; Experimental Assay: Cm ionic:LiBr: 1.52 M ; Experimental Assay: m ionic:LiBr: 1.52 M ; Experimental Assay: dG_H2O ionic:LiBr: 1.52 M ; Experimental Assay: Cm ionic:LiBr: 0.76 M ; Experimental Assay: m ionic:LiBr: 0.76 M ; Experimental Assay: dG_H2O ionic:LiBr: 0.76 M ; Experimental Assay: Cm ionic:NaBr: 0.99 M ; Experimental Assay: m ionic:NaBr: 0.99 M ; Experimental Assay: dG_H2O ionic:NaBr: 0.99 M ; Experimental Assay: Cm ionic:NaBr: 0.50 M ; Experimental Assay: m ionic:NaBr: 0.50 M ; Experimental Assay: dG_H2O ionic:NaBr: 0.50 M ; Experimental Assay: Cm ionic:Li2SO4: 0.40 M ; Experimental Assay: m ionic:Li2SO4: 0.40 M ; Experimental Assay: dG_H2O ionic:Li2SO4: 0.40 M ; Experimental Assay: Cm ionic:Li2SO4: 0.20 M ; Experimental Assay: m ionic:Li2SO4: 0.20 M ; Experimental Assay: dG_H2O ionic:Li2SO4: 0.20 M ; Experimental Assay: Cm ionic:(NH4)2SO4: 0.30 M ; Experimental Assay: m ionic:(NH4)2SO4: 0.30 M ; Experimental Assay: dG_H2O ionic:(NH4)2SO4: 0.30 M ; Experimental Assay: Cm ionic:(NH4)2SO4: 0.15 M ; Experimental Assay: m ionic:(NH4)2SO4: 0.15 M ; Experimental Assay: dG_H2O ionic:(NH4)2SO4: 0.15 M ; Experimental Assay: Cm ionic:Na2SO4: 0.30 M ; Experimental Assay: m ionic:Na2SO4: 0.30 M ; Experimental Assay: dG_H2O ionic:Na2SO4: 0.30 M ; Experimental Assay: Cm ionic:Na2SO4: 0.15 M ; Experimental Assay: m ionic:Na2SO4: 0.15 M ; Experimental Assay: dG_H2O ionic:Na2SO4: 0.15 M ; Experimental Assay: Cm ionic:LiCi: 0.99 M ; Experimental Assay: m ionic:LiCi: 0.99 M ; Experimental Assay: dG_H2O ionic:LiCi: 0.99 M ; Experimental Assay: Cm ionic:LiCi: 0.50 M ; Experimental Assay: m ionic:LiCi: 0.50 M ; Experimental Assay: dG_H2O ionic:LiCi: 0.50 M ; Experimental Assay: Cm ionic:NH4Cl: 0.45 M ; Experimental Assay: m ionic:NH4Cl: 0.45 M ; Experimental Assay: dG_H2O ionic:NH4Cl: 0.45 M ; Experimental Assay: Cm ionic:RbCl: 0.50 M ; Experimental Assay: m ionic:RbCl: 0.50 M ; Experimental Assay: dG_H2O ionic:RbCl: 0.50 M ; Experimental Assay: Cm ionic:CsCl: 0.49 M ; Experimental Assay: m ionic:CsCl: 0.49 M ; Experimental Assay: dG_H2O ionic:CsCl: 0.49 M ; Experimental Assay: Cm ionic:KCl: 0.1 M ; Experimental Assay: m ionic:KCl: 0.1 M ; Experimental Assay: dG_H2O ionic:KCl: 0.1 M ; Experimental Assay: Cm ionic:NaCl: 0.1 M ; Experimental Assay: m ionic:NaCl: 0.1 M ; Experimental Assay: dG_H2O ionic:NaCl: 0.1 M
Libraries Mutations for sequence KETAAAKFERQHMDSSTSAASSSNYCNQMMKSRNLTKDRCKPVNTFVHESLADVQAVCSQKNVACKNGQTNCYQSYSTMSITDCRETGSSKYPNCAYKTTQANKHIIVACEGNPYVPVHFDASV

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
6QMN 2019-02-07T00:00:00+0000 2.31 Crystal structure of a Ribonuclease A-Onconase chimera
1A2W 1998-01-12T00:00:00+0000 2.1 CRYSTAL STRUCTURE OF A 3D DOMAIN-SWAPPED DIMER OF BOVINE PANCREATIC RIBONUCLEASE A
1A5P 1998-02-17T00:00:00+0000 1.6 C[40,95]A VARIANT OF BOVINE PANCREATIC RIBONUCLEASE A
1A5Q 1998-02-17T00:00:00+0000 2.3 P93A VARIANT OF BOVINE PANCREATIC RIBONUCLEASE A
1AFK 1997-03-08T00:00:00+0000 1.7 CRYSTAL STRUCTURE OF RIBONUCLEASE A IN COMPLEX WITH 5'-DIPHOSPHOADENOSINE-3'-PHOSPHATE
1AFL 1997-03-08T00:00:00+0000 1.7 RIBONUCLEASE A IN COMPLEX WITH 5'-DIPHOSPHOADENOSINE 2'-PHOSPHATE AT 1.7 ANGSTROM RESOLUTION
1AFU 1997-03-14T00:00:00+0000 2.0 STRUCTURE OF RIBONUCLEASE A AT 2.0 ANGSTROMS FROM MONOCLINIC CRYSTALS
1AQP 1997-07-31T00:00:00+0000 2.0 RIBONUCLEASE A COPPER COMPLEX
1B6V 1999-01-18T00:00:00+0000 2.0 CRYSTAL STRUCTURE OF A HYBRID BETWEEN RIBONUCLEASE A AND BOVINE SEMINAL RIBONUCLEASE
1BEL 1995-12-21T00:00:00+0000 1.6 HYDROLASE PHOSPHORIC DIESTER, RNA

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Ribonuclease pancreatic P61824 RNAS1_BISBI
100.0 Ribonuclease pancreatic P61823 RNAS1_BOVIN
96.8 Ribonuclease pancreatic P67926 RNAS1_CAPHI
96.8 Ribonuclease pancreatic P67927 RNAS1_SHEEP
95.2 Ribonuclease pancreatic P00657 RNAS1_BUBBU
96.0 Ribonuclease pancreatic P07847 RNAS1_AEPME
93.5 Ribonuclease pancreatic P07848 RNAS1_EUDTH
95.2 Ribonuclease pancreatic P00660 RNAS1_CONTA
92.7 Ribonuclease pancreatic P00668 RNAS1_ANTAM
90.3 Ribonuclease pancreatic P00662 RNAS1_GIRCA
96.0 Ribonuclease pancreatic Q29606 RNAS1_ORYLE