Determining stability of proteins from guanidinium chloride transition curves.


Abstract

The guanidinium chloride (GdmCl) denaturation of RNAase A, lysozyme and metmyoglobin was investigated at several pH values by using absorbance measurements at 287, 300 and 409 nm respectively. From these measurements the free-energy change on denaturation, delta Gapp., was calculated, assuming a two-state mechanism, and values of delta Gapp. at zero concentration of the denaturant were measured. For each protein all delta Gapp. values were adjusted to pH 7.00 by using the appropriate relationship between delta Gapp. and pH. Dependence of the adjusted delta Gapp. value on GdmCl concentration increases for metmyoglobin and decreases for the other two proteins as the denaturant concentration decreases. It has been shown that these are expected results if the presence of the acid-denatured state during the GdmCl denaturation of proteins is considered. Study holds ProTherm entries: 7718, 7719, 7720, 7721, 7722, 7723, 7724, 7725, 7726, 7727, 7728, 7729, 7730, 7731, 7732 Extra Details: guanidinium chloride transition curve; absorbance measurement;,free-energy change; two-state mechanism; acid-denatured state

Submission Details

ID: mX63tSLC3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:34 p.m.

Version: 1

Publication Details
Ahmad F;Yadav S;Taneja S,Biochem. J. (1992) Determining stability of proteins from guanidinium chloride transition curves. PMID:1445206
Additional Information

Number of data points 45
Proteins Ribonuclease pancreatic ; Myoglobin ; Lysozyme C ; Ribonuclease pancreatic ; Myoglobin ; Lysozyme C
Unique complexes 3
Assays/Quantities/Protocols Experimental Assay: Cm pH:4.6 ; Experimental Assay: m pH:4.6 ; Experimental Assay: dG_H2O pH:4.6 ; Experimental Assay: Cm pH:4.92 ; Experimental Assay: m pH:4.92 ; Experimental Assay: dG_H2O pH:4.92 ; Experimental Assay: Cm pH:5.26 ; Experimental Assay: m pH:5.26 ; Experimental Assay: dG_H2O pH:5.26 ; Experimental Assay: Cm pH:5.46 ; Experimental Assay: m pH:5.46 ; Experimental Assay: dG_H2O pH:5.46 ; Experimental Assay: Cm pH:5.52 ; Experimental Assay: m pH:5.52 ; Experimental Assay: dG_H2O pH:5.52 ; Experimental Assay: Cm pH:5.88 ; Experimental Assay: m pH:5.88 ; Experimental Assay: dG_H2O pH:5.88 ; Experimental Assay: Cm pH:0.34 ; Experimental Assay: m pH:0.34 ; Experimental Assay: dG_H2O pH:0.34 ; Experimental Assay: Cm pH:0.64 ; Experimental Assay: m pH:0.64 ; Experimental Assay: dG_H2O pH:0.64 ; Experimental Assay: Cm pH:2.03 ; Experimental Assay: m pH:2.03 ; Experimental Assay: dG_H2O pH:2.03 ; Experimental Assay: Cm pH:6.0 ; Experimental Assay: m pH:6.0 ; Experimental Assay: dG_H2O pH:6.0 ; Experimental Assay: Cm pH:1.3 ; Experimental Assay: m pH:1.3 ; Experimental Assay: dG_H2O pH:1.3 ; Experimental Assay: Cm pH:2.1 ; Experimental Assay: m pH:2.1 ; Experimental Assay: dG_H2O pH:2.1 ; Experimental Assay: Cm pH:3.0 ; Experimental Assay: m pH:3.0 ; Experimental Assay: dG_H2O pH:3.0 ; Experimental Assay: Cm pH:7.0 ; Experimental Assay: m pH:7.0 ; Experimental Assay: dG_H2O pH:7.0
Libraries Mutations for sequence KETAAAKFERQHMDSSTSAASSSNYCNQMMKSRNLTKDRCKPVNTFVHESLADVQAVCSQKNVACKNGQTNCYQSYSTMSITDCRETGSSKYPNCAYKTTQANKHIIVACEGNPYVPVHFDASV ; Mutations for sequence GLSDGEWQQVLNVWGKVEADIAGHGQEVLIRLFTGHPETLEKFDKFKHLKTEAEMKASEDLKKHGTVVLTALGGILKKKGHHEAELKPLAQSHATKHKIPIKYLEFISDAIIHVLHSKHPGDFGADAQGAMTKALELFRNDIAAKYKELGFQG ; Mutations for sequence KVFGRCELAAAMKRHGLDNYRGYSLGNWVCAAKFESNFNTQATNRNTDGSTDYGILQINSRWWCNDGRTPGSRNLCNIPCSALLSSDITASVNCAKKIVSDGNGMNAWVAWRNRCKGTDVQAWIRGCRL
Sequence Assay Result Units