A new method for determining the heat capacity change for protein folding.


Abstract

In order to use results from calorimetry or thermal unfolding curves to estimate the free energy change for protein unfolding at 25 degrees C, it is necessary to know the change in heat capacity for unfolding, delta Cp. We describe a new method for measuring delta Cp which is based on results from urea and thermal unfolding curves but does not require a calorimeter. We find that delta Cp = 1650 +/- 200 cal/(deg.mol) for the unfolding of ribonuclease T1 and that delta Cp = 2200 +/- 300 cal/(deg.mol) for the unfolding of ribonuclease A. Study holds ProTherm entries: 3780, 3781, 3782, 3783, 3784, 3785, 3786, 3787, 3788, 3789, 3790, 3791, 3792, 3793, 3794, 3795, 3796, 3797, 3798, 3799, 3800, 3801 Extra Details: heat capacity change; protein folding;,thermal unfolding curves; free energy change

Submission Details

ID: p4fK6s8c

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:22 p.m.

Version: 1

Publication Details
Pace CN;Laurents DV,Biochemistry (1989) A new method for determining the heat capacity change for protein folding. PMID:2499351
Additional Information

Study Summary

Number of data points 84
Proteins Guanyl-specific ribonuclease T1 ; Guanyl-specific ribonuclease T1
Unique complexes 2
Assays/Quantities/Protocols Experimental Assay: Cm temp:27.75 C, buffers:glycine: 30 mM, pH:2.8 ; Experimental Assay: dCp temp:27.75 C, buffers:glycine: 30 mM, pH:2.8 ; Experimental Assay: m temp:27.75 C, buffers:glycine: 30 mM, pH:2.8 ; Experimental Assay: dG_H2O temp:27.75 C, buffers:glycine: 30 mM, pH:2.8 ; Experimental Assay: Cm temp:24.9 C, buffers:glycine: 30 mM, pH:2.8 ; Experimental Assay: dCp temp:24.9 C, buffers:glycine: 30 mM, pH:2.8 ; Experimental Assay: m temp:24.9 C, buffers:glycine: 30 mM, pH:2.8 ; Experimental Assay: dG_H2O temp:24.9 C, buffers:glycine: 30 mM, pH:2.8 ; Experimental Assay: Cm buffers:glycine: 30 mM, pH:2.8, temp:21.1 C ; Experimental Assay: dCp buffers:glycine: 30 mM, pH:2.8, temp:21.1 C ; Experimental Assay: m buffers:glycine: 30 mM, pH:2.8, temp:21.1 C ; Experimental Assay: dG_H2O buffers:glycine: 30 mM, pH:2.8, temp:21.1 C ; Experimental Assay: Cm temp:17.1 C, pH:2.8, buffers:glycine: 30 mM ; Experimental Assay: dCp temp:17.1 C, pH:2.8, buffers:glycine: 30 mM ; Experimental Assay: m temp:17.1 C, pH:2.8, buffers:glycine: 30 mM ; Experimental Assay: dG_H2O temp:17.1 C, pH:2.8, buffers:glycine: 30 mM ; Experimental Assay: Cm buffers:HEPPSO: 30 mM, pH:7.9, temp:28.95 C ; Experimental Assay: dCp buffers:HEPPSO: 30 mM, pH:7.9, temp:28.95 C ; Experimental Assay: m buffers:HEPPSO: 30 mM, pH:7.9, temp:28.95 C ; Experimental Assay: dG_H2O buffers:HEPPSO: 30 mM, pH:7.9, temp:28.95 C ; Experimental Assay: Cm buffers:HEPPSO: 30 mM, pH:7.9, temp:25.0 C ; Experimental Assay: dCp buffers:HEPPSO: 30 mM, pH:7.9, temp:25.0 C ; Experimental Assay: m buffers:HEPPSO: 30 mM, pH:7.9, temp:25.0 C ; Experimental Assay: dG_H2O buffers:HEPPSO: 30 mM, pH:7.9, temp:25.0 C ; Experimental Assay: Cm temp:22.0 C, buffers:HEPPSO: 30 mM, pH:7.9 ; Experimental Assay: dCp temp:22.0 C, buffers:HEPPSO: 30 mM, pH:7.9 ; Experimental Assay: m temp:22.0 C, buffers:HEPPSO: 30 mM, pH:7.9 ; Experimental Assay: dG_H2O temp:22.0 C, buffers:HEPPSO: 30 mM, pH:7.9 ; Experimental Assay: Cm buffers:HEPPSO: 30 mM, temp:19.05 C, pH:7.9 ; Experimental Assay: dCp buffers:HEPPSO: 30 mM, temp:19.05 C, pH:7.9 ; Experimental Assay: m buffers:HEPPSO: 30 mM, temp:19.05 C, pH:7.9 ; Experimental Assay: dG_H2O buffers:HEPPSO: 30 mM, temp:19.05 C, pH:7.9 ; Experimental Assay: Cm temp:29.0 C, buffers:MOPS: 30 mM, pH:7.0 ; Experimental Assay: dCp temp:29.0 C, buffers:MOPS: 30 mM, pH:7.0 ; Experimental Assay: m temp:29.0 C, buffers:MOPS: 30 mM, pH:7.0 ; Experimental Assay: dG_H2O temp:29.0 C, buffers:MOPS: 30 mM, pH:7.0 ; Experimental Assay: Cm buffers:MOPS: 30 mM, temp:27.0 C, pH:7.0 ; Experimental Assay: dCp pH:7.0, buffers:MOPS: 30 mM, temp:27.0 C ; Experimental Assay: m buffers:MOPS: 30 mM, temp:27.0 C, pH:7.0 ; Experimental Assay: dG_H2O buffers:MOPS: 30 mM, temp:27.0 C, pH:7.0 ; Experimental Assay: Cm buffers:MOPS: 30 mM, temp:25.0 C, pH:7.0 ; Experimental Assay: dCp buffers:MOPS: 30 mM, pH:7.0, temp:25.0 C ; Experimental Assay: m buffers:MOPS: 30 mM, temp:25.0 C, pH:7.0 ; Experimental Assay: dG_H2O buffers:MOPS: 30 mM, temp:25.0 C, pH:7.0 ; Experimental Assay: Cm buffers:MOPS: 30 mM, temp:23.05 C, pH:7.0 ; Experimental Assay: dCp buffers:MOPS: 30 mM, pH:7.0, temp:23.05 C ; Experimental Assay: m buffers:MOPS: 30 mM, temp:23.05 C, pH:7.0 ; Experimental Assay: dG_H2O buffers:MOPS: 30 mM, temp:23.05 C, pH:7.0 ; Experimental Assay: Cm buffers:MOPS: 30 mM, temp:21.1 C, pH:7.0 ; Experimental Assay: dCp buffers:MOPS: 30 mM, pH:7.0, temp:21.1 C ; Experimental Assay: m buffers:MOPS: 30 mM, temp:21.1 C, pH:7.0 ; Experimental Assay: dG_H2O buffers:MOPS: 30 mM, temp:21.1 C, pH:7.0 ; Experimental Assay: Cm buffers:MOPS: 30 mM, temp:19.2 C, pH:7.0 ; Experimental Assay: dCp buffers:MOPS: 30 mM, pH:7.0, temp:19.2 C ; Experimental Assay: m buffers:MOPS: 30 mM, temp:19.2 C, pH:7.0 ; Experimental Assay: dG_H2O buffers:MOPS: 30 mM, temp:19.2 C, pH:7.0 ; Experimental Assay: Cm pH:5.95, buffers:MES: 30 mM, temp:29.1 C ; Experimental Assay: dCp pH:5.95, buffers:MES: 30 mM, temp:29.1 C ; Experimental Assay: m pH:5.95, buffers:MES: 30 mM, temp:29.1 C ; Experimental Assay: dG_H2O pH:5.95, buffers:MES: 30 mM, temp:29.1 C ; Experimental Assay: Cm pH:5.95, buffers:MES: 30 mM, temp:25.05 C ; Experimental Assay: dCp pH:5.95, buffers:MES: 30 mM, temp:25.05 C ; Experimental Assay: m pH:5.95, buffers:MES: 30 mM, temp:25.05 C ; Experimental Assay: dG_H2O pH:5.95, buffers:MES: 30 mM, temp:25.05 C ; Experimental Assay: Cm pH:5.95, temp:22.0 C, buffers:MES: 30 mM ; Experimental Assay: dCp pH:5.95, temp:22.0 C, buffers:MES: 30 mM ; Experimental Assay: m pH:5.95, temp:22.0 C, buffers:MES: 30 mM ; Experimental Assay: dG_H2O pH:5.95, temp:22.0 C, buffers:MES: 30 mM ; Experimental Assay: Cm pH:5.95, temp:19.05 C, buffers:MES: 30 mM ; Experimental Assay: dCp pH:5.95, temp:19.05 C, buffers:MES: 30 mM ; Experimental Assay: m pH:5.95, temp:19.05 C, buffers:MES: 30 mM ; Experimental Assay: dG_H2O pH:5.95, temp:19.05 C, buffers:MES: 30 mM ; Experimental Assay: dCp pH:2.8, buffers:glycine: 30 mM ; Experimental Assay: dHcal buffers:glycine: 30 mM, pH:2.8 ; Experimental Assay: Tm buffers:glycine: 30 mM, pH:2.8 ; Experimental Assay: dCp buffers:HEPPSO: 30 mM, pH:7.9 ; Experimental Assay: dHcal buffers:HEPPSO: 30 mM, pH:7.9 ; Experimental Assay: Tm buffers:HEPPSO: 30 mM, pH:7.9 ; Experimental Assay: dCp buffers:MOPS: 30 mM, pH:7.0 ; Experimental Assay: dHcal buffers:MOPS: 30 mM, pH:7.0 ; Experimental Assay: Tm buffers:MOPS: 30 mM, pH:7.0 ; Experimental Assay: dCp pH:5.95, buffers:MES: 30 mM ; Experimental Assay: dHcal pH:5.95, buffers:MES: 30 mM ; Experimental Assay: Tm pH:5.95, buffers:MES: 30 mM
Libraries Mutations for sequence MMYSKLLTLTTLLLPTALALPSLVERACDYTCGSNCYSSSDVSTAQAAGYQLHEDGETVGSNSYPHKYNNYEGFDFSVSSPYYEWPILSSGDVYSGGSPGADRVVFNENNQLAGVITHTGASGNNFVECT ; Mutations for sequence ACDYTCGSNCYSSSDVSTAQAAGYQLHEDGETVGSNSYPHKYNNYEGFDFSVSSPYYEWPILSSGDVYSGGSPGADRVVFNENNQLAGVITHTGASGNNFVECT

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
1B2M 1998-11-27T00:00:00+0000 2.0 THREE-DIMENSIONAL STRUCTURE OF RIBONULCEASE T1 COMPLEXED WITH AN ISOSTERIC PHOSPHONATE ANALOGUE OF GPU: ALTERNATE SUBSTRATE BINDING MODES AND CATALYSIS.
1BIR 1996-01-04T00:00:00+0000 1.8 RIBONUCLEASE T1, PHE 100 TO ALA MUTANT COMPLEXED WITH 2' GMP
1BU4 1998-09-11T00:00:00+0000 1.9 RIBONUCLEASE 1 COMPLEX WITH 2'GMP
1BVI 1998-09-15T00:00:00+0000 1.9 RIBONUCLEASE T1 (WILDTYPE) COMPLEXED WITH 2'GMP
1CH0 1999-03-30T00:00:00+0000 2.3 RNASE T1 VARIANT WITH ALTERED GUANINE BINDING SEGMENT
1DET 1996-02-20T00:00:00+0000 1.8 RIBONUCLEASE T1 CARBOXYMETHYLATED AT GLU 58 IN COMPLEX WITH 2'GMP
1FYS 2000-10-03T00:00:00+0000 2.0 Ribonuclease T1 V16C mutant
1FZU 2000-10-04T00:00:00+0000 1.8 RNAse T1 V78A mutant
1G02 2000-10-05T00:00:00+0000 1.86 Ribonuclease T1 V16S mutant
1GSP 1997-11-28T00:00:00+0000 2.2 RIBONUCLEASE T1 COMPLEXED WITH 2',3'-CGPS, 1 DAY

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Guanyl-specific ribonuclease T1 P00651 RNT1_ASPOR