Thermodynamics of the thermal unfolding of eglin c in the presence and absence of guanidinium chloride.


Abstract

The thermal unfolding of eglin c, a small proteinase inhibitor of molecular weight 8.1 kDa, is studied by means of high sensitivity scanning calorimetry over a wide pH range in dilute buffer solutions, and in the presence of varying concentrations of guanidinium chloride at pH 7.00 and 10.55. The temperature of half-completion of the unfolding transition, t1/2, in dilute buffer varies from 41 degrees C at pH 1.1 to 86 degrees C at pH 7.0 to 10.55, with corresponding enthalpy changes of approximately 40 kcal mol-1 and 71 kcal mol-1. This latter enthalpy change, amounting to 8.7 cal g-1, is unusually large for a protein, especially for one of unusually small molecular weight. Addition of 3.3 M guanidinium chloride at pH 10.55 lowered t1/2 from 86 degrees C to 40 degrees C and decreased the enthalpy change from approximately 71 kcal mol-1 to 25 kcal mol-1. Study holds ProTherm entries: 9898, 9899, 9900, 9901, 9902, 9903, 9904, 9905, 9906, 9907, 9908, 9909, 9910, 9911, 9912, 9913, 9914, 9915, 9916, 9917, 9918 Extra Details: thermodynamics; thermal unfolding; eglin c; guanidinium chloride

Submission Details

ID: W4up4FzT4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:39 p.m.

Version: 1

Publication Details
Bae SJ;Sturtevant JM,Biophys. Chem. (1995) Thermodynamics of the thermal unfolding of eglin c in the presence and absence of guanidinium chloride. PMID:7626743
Additional Information

Study Summary

Number of data points 63
Proteins Chymotrypsinogen A ; Eglin C
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: dCp buffers:glycine: 50mM, pH:10.55 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:10.55 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:10.55 ; Experimental Assay: dCp buffers:glycine: 50mM, pH:10.5 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:10.5 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:10.5 ; Experimental Assay: dCp buffers:glycine: 50mM, pH:10.05 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:10.05 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:10.05 ; Experimental Assay: dCp pH:9.2, buffers:bicine: 50mM ; Experimental Assay: dHcal pH:9.2, buffers:bicine: 50mM ; Experimental Assay: Tm pH:9.2, buffers:bicine: 50mM ; Experimental Assay: dCp buffers:Sodium phosphate: 50mM, pH:8.0 ; Experimental Assay: dHcal buffers:Sodium phosphate: 50mM, pH:8.0 ; Experimental Assay: Tm buffers:Sodium phosphate: 50mM, pH:8.0 ; Experimental Assay: dCp buffers:Sodium phosphate: 50mM, pH:7.05 ; Experimental Assay: dHcal buffers:Sodium phosphate: 50mM, pH:7.05 ; Experimental Assay: Tm buffers:Sodium phosphate: 50mM, pH:7.05 ; Experimental Assay: dCp pH:7.01, buffers:Sodium phosphate: 50mM ; Experimental Assay: dHcal pH:7.01, buffers:Sodium phosphate: 50mM ; Experimental Assay: Tm pH:7.01, buffers:Sodium phosphate: 50mM ; Experimental Assay: dCp buffers:Sodium phosphate: 50mM, pH:7.0 ; Experimental Assay: dHcal buffers:Sodium phosphate: 50mM, pH:7.0 ; Experimental Assay: Tm buffers:Sodium phosphate: 50mM, pH:7.0 ; Experimental Assay: dCp buffers:glycine: 50mM, pH:3.25 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:3.25 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:3.25 ; Experimental Assay: dCp buffers:glycine: 50mM, pH:3.0 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:3.0 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:3.0 ; Experimental Assay: dCp buffers:glycine: 50mM, pH:2.93 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:2.93 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:2.93 ; Experimental Assay: dCp buffers:glycine: 50mM, pH:2.85 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:2.85 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:2.85 ; Experimental Assay: dCp buffers:glycine: 50mM, pH:2.75 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:2.75 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:2.75 ; Experimental Assay: dCp buffers:glycine: 50mM, pH:2.5 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:2.5 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:2.5 ; Experimental Assay: dCp buffers:glycine: 50mM, pH:2.25 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:2.25 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:2.25 ; Experimental Assay: dCp buffers:glycine: 50mM, pH:2.15 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:2.15 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:2.15 ; Experimental Assay: dCp buffers:glycine: 50mM, pH:2.0 ; Experimental Assay: dHcal buffers:glycine: 50mM, pH:2.0 ; Experimental Assay: Tm buffers:glycine: 50mM, pH:2.0 ; Experimental Assay: dCp pH:1.8, buffers:glycine-HCl: 50mM ; Experimental Assay: dHcal pH:1.8, buffers:glycine-HCl: 50mM ; Experimental Assay: Tm pH:1.8, buffers:glycine-HCl: 50mM ; Experimental Assay: dCp buffers:glycine-HCl: 50mM, pH:1.65 ; Experimental Assay: dHcal buffers:glycine-HCl: 50mM, pH:1.65 ; Experimental Assay: Tm buffers:glycine-HCl: 50mM, pH:1.65 ; Experimental Assay: dCp pH:1.3, buffers:glycine-HCl: 50mM ; Experimental Assay: dHcal pH:1.3, buffers:glycine-HCl: 50mM ; Experimental Assay: Tm pH:1.3, buffers:glycine-HCl: 50mM ; Experimental Assay: dCp pH:1.1, buffers:glycine-HCl: 50mM ; Experimental Assay: dHcal pH:1.1, buffers:glycine-HCl: 50mM ; Experimental Assay: Tm pH:1.1, buffers:glycine-HCl: 50mM
Libraries Mutations for sequence E:CGVPAIQPVLSGLSRIVNGEEAVPGSWPWQVSLQDKTGFHFCGGSLINENWVVTAAHCGVTTSDVVVAGEFDQGSSSEKIQKLKIAKVFKNSKYNSLTINNDITLLKLSTAASFSQTVSAVCLPSASDDFAAGTTCVTTGWGLTRYTNANTPDRLQQASLPLLSNTNCKKYWGTKIKDAMICAGASGVSSCMGDSGGPLVCKKNGAWTLVGIVSWGSSTCSTSTPGVYARVTALVNWVQQTLAAN/I:TEFGSELKSFPEVVGKTVDQAREYFTLHYPQYDVYFLPEGSPVTLDLRYNRVRVFYNPGTNVVNHVPHVG

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
1EGL 1994-01-31 THE SOLUTION STRUCTURE OF EGLIN C BASED ON MEASUREMENTS OF MANY NOES AND COUPLING CONSTANTS AND ITS COMPARISON WITH X-RAY STRUCTURES
1CSE 1988-07-16 1.2 THE HIGH-RESOLUTION X-RAY CRYSTAL STRUCTURE OF THE COMPLEX FORMED BETWEEN SUBTILISIN CARLSBERG AND EGLIN C, AN ELASTASE INHIBITOR FROM THE LEECH HIRUDO MEDICINALIS. STRUCTURAL ANALYSIS, SUBTILISIN STRUCTURE AND INTERFACE GEOMETRY
1YPH 2006-02-14 1.34 High resolution structure of bovine alpha-chymotrypsin
4B2B 2012-08-01 1.36 Structure of the factor Xa-like trypsin variant triple-Ala (TGPA) in complex with eglin C
1GG6 2000-09-20 1.4 CRYSTAL STRUCTURE OF GAMMA CHYMOTRYPSIN WITH N-ACETYL-PHENYLALANINE TRIFLUOROMETHYL KETONE BOUND AT THE ACTIVE SITE
4B2C 2012-08-01 1.43 Structure of the factor Xa-like trypsin variant triple-Ala (TPA) in complex with eglin C
2P8O 2007-05-08 1.5 Crystal Structure of a Benzohydroxamic Acid/Vanadate complex bound to chymotrypsin A
1GGD 2000-09-20 1.5 CRYSTAL STRUCTURE OF GAMMA CHYMOTRYPSIN WITH N-ACETYL-LEUCIL-PHENYLALANINE ALDEHYDE BOUND AT THE ACTIVE SITE
1AB9 1997-08-20 1.6 CRYSTAL STRUCTURE OF BOVINE GAMMA-CHYMOTRYPSIN
1GCT 1991-10-15 1.6 IS GAMMA-CHYMOTRYPSIN A TETRAPEPTIDE ACYL-ENZYME ADDUCT OF GAMMA-CHYMOTRYPSIN?
8GCH 1992-04-15 1.6 GAMMA-CHYMOTRYPSIN IS A COMPLEX OF ALPHA-CHYMOTRYPSIN WITH ITS OWN AUTOLYSIS PRODUCTS
3GCT 1991-10-15 1.6 STRUCTURE OF GAMMA-*CHYMOTRYPSIN IN THE RANGE $P*H 2.0 TO $P*H 10.5 SUGGESTS THAT GAMMA-CHYMOTRYPSIN IS A COVALENT ACYL-ENZYME ADDUCT AT LOW $P*H
3VGC 1997-11-12 1.67 GAMMA-CHYMOTRYPSIN L-NAPHTHYL-1-ACETAMIDO BORONIC ACID ACID INHIBITOR COMPLEX
5CHA 1985-04-01 1.67 THE REFINEMENT AND THE STRUCTURE OF THE DIMER OF ALPHA-*CHYMOTRYPSIN AT 1.67-*ANGSTROMS RESOLUTION
3RU4 2012-08-29 1.68 Crystal structure of the Bowman-Birk serine protease inhibitor BTCI in complex with trypsin and chymotrypsin
4CHA 1985-04-01 1.68 STRUCTURE OF ALPHA-*CHYMOTRYPSIN REFINED AT 1.68 ANGSTROMS RESOLUTION
1T8O 2005-03-08 1.7 CRYSTAL STRUCTURE OF THE P1 TRP BPTI MUTANT- BOVINE CHYMOTRYPSIN COMPLEX
1T8L 2005-03-08 1.75 CRYSTAL STRUCTURE OF THE P1 MET BPTI MUTANT- BOVINE CHYMOTRYPSIN COMPLEX
1P2M 2004-04-20 1.75 Structural consequences of accommodation of four non-cognate amino-acid residues in the S1 pocket of bovine trypsin and chymotrypsin
1T8N 2005-03-08 1.75 CRYSTAL STRUCTURE OF THE P1 THR BPTI MUTANT- BOVINE CHYMOTRYPSIN COMPLEX
4B1T 2012-08-01 1.78 Structure of the factor Xa-like trypsin variant triple-Ala (TA) in complex with eglin C
1T8M 2005-03-08 1.8 CRYSTAL STRUCTURE OF THE P1 HIS BPTI MUTANT- BOVINE CHYMOTRYPSIN COMPLEX
2CGA 1987-04-16 1.8 BOVINE CHYMOTRYPSINOGEN A. X-RAY CRYSTAL STRUCTURE ANALYSIS AND REFINEMENT OF A NEW CRYSTAL FORM AT 1.8 ANGSTROMS RESOLUTION
1AFQ 1997-09-17 1.8 CRYSTAL STRUCTURE OF BOVINE GAMMA-CHYMOTRYPSIN COMPLEXED WITH A SYNTHETIC INHIBITOR
1P2N 2004-04-20 1.8 Structural consequences of accommodation of four non-cognate amino-acid residues in the S1 pocket of bovine trypsin and chymotrypsin
2SEC 1988-09-07 1.8 STRUCTURAL COMPARISON OF TWO SERINE PROTEINASE-PROTEIN INHIBITOR COMPLEXES. EGLIN-C-SUBTILISIN CARLSBERG AND CI-2-SUBTILISIN NOVO
7GCH 1990-10-15 1.8 STRUCTURE OF CHYMOTRYPSIN-*TRIFLUOROMETHYL KETONE INHIBITOR COMPLEXES. COMPARISON OF SLOWLY AND RAPIDLY EQUILIBRATING INHIBITORS
2GMT 1994-11-01 1.8 THREE-DIMENSIONAL STRUCTURE OF CHYMOTRYPSIN INACTIVATED WITH (2S) N-ACETYL-L-ALANYL-L-PHENYLALANYL-CHLOROETHYL KETONE: IMPLICATIONS FOR THE MECHANISM OF INACTIVATION OF SERINE PROTEASES BY CHLOROKETONES
1CHO 1988-07-16 1.8 CRYSTAL AND MOLECULAR STRUCTURES OF THE COMPLEX OF ALPHA-*CHYMOTRYPSIN WITH ITS INHIBITOR TURKEY OVOMUCOID THIRD DOMAIN AT 1.8 ANGSTROMS RESOLUTION
1P2Q 2004-04-20 1.8 Structural consequences of accommodation of four non-cognate amino-acid residues in the S1 pocket of bovine trypsin and chymotrypsin
1K2I 2001-12-05 1.8 Crystal Structure of Gamma-Chymotrypsin in Complex with 7-Hydroxycoumarin
2GCT 1991-10-15 1.8 STRUCTURE OF GAMMA-CHYMOTRYPSIN IN THE RANGE PH 2.0 TO PH 10.5 SUGGESTS THAT GAMMA-CHYMOTRYPSIN IS A COVALENT ACYL-ENZYME ADDUCT AT LOW PH
2VGC 1997-11-12 1.8 GAMMA-CHYMOTRYPSIN D-PARA-CHLORO-1-ACETAMIDO BORONIC ACID INHIBITOR COMPLEX
6CHA 1987-04-16 1.8 STRUCTURE OF A TETRAHEDRAL TRANSITION STATE COMPLEX OF ALPHA-*CHYMOTRYPSIN AT 1.8-*ANGSTROMS RESOLUTION
1T7C 2005-03-08 1.85 CRYSTAL STRUCTURE OF THE P1 GLU BPTI MUTANT- BOVINE CHYMOTRYPSIN COMPLEX
6DI8 2019-01-23 1.86 Crystal structure of bovine alpha-chymotrypsin in space group P65
4B2A 2012-08-01 1.89 Structure of the factor Xa-like trypsin variant triple-Ala (TGA) in complex with eglin C
4GCH 1990-10-15 1.9 STRUCTURE AND ACTIVITY OF TWO PHOTOREVERSIBLE CINNAMATES BOUND TO CHYMOTRYPSIN
1VGC 1997-11-12 1.9 GAMMA-CHYMOTRYPSIN L-PARA-CHLORO-1-ACETAMIDO BORONIC ACID INHIBITOR COMPLEX
2GCH 1980-07-09 1.9 REFINED CRYSTAL STRUCTURE OF GAMMA-CHYMOTRYPSIN AT 1.9 ANGSTROMS RESOLUTION
4H4F 2013-02-27 1.9 Crystal structure of human chymotrypsin C (CTRC) bound to inhibitor eglin c from Hirudo medicinalis
1GCD 1994-06-22 1.9 REFINED CRYSTAL STRUCTURE OF 'AGED' AND 'NON-AGED' ORGANOPHOSPHORYL CONJUGATES OF GAMMA-CHYMOTRYPSIN
3GCH 1990-10-15 1.9 CHEMISTRY OF CAGED ENZYMES. BINDING OF PHOTOREVERSIBLE CINNAMATES TO CHYMOTRYPSIN
2TEC 1992-01-15 1.98 MOLECULAR DYNAMICS REFINEMENT OF A THERMITASE-EGLIN-C COMPLEX AT 1.98 ANGSTROMS RESOLUTION AND COMPARISON OF TWO CRYSTAL FORMS THAT DIFFER IN CALCIUM CONTENT
1GHB 1994-06-22 2.0 A SECOND ACTIVE SITE IN CHYMOTRYPSIN? THE X-RAY CRYSTAL STRUCTURE OF N-ACETYL-D-TRYPTOPHAN BOUND TO GAMMA-CHYMOTRYPSIN
2CHA 1977-01-18 2.0 THE STRUCTURE OF CRYSTALLINE ALPHA-CHYMOTRYPSIN, $V.THE ATOMIC STRUCTURE OF TOSYL-ALPHA-CHYMOTRYPSIN AT 2 ANGSTROMS RESOLUTION
3T62 2012-08-01 2.0 Crystal structure of recombinant Kunitz Type serine protease Inhibitor-1 from the Caribbean Sea anemone Stichodactyla helianthus in complex with bovine chymotrypsin
3TEC 1992-01-15 2.0 CALCIUM BINDING TO THERMITASE. CRYSTALLOGRAPHIC STUDIES OF THERMITASE AT 0, 5 AND 100 MM CALCIUM
1ACB 1993-10-31 2.0 CRYSTAL AND MOLECULAR STRUCTURE OF THE BOVINE ALPHA-CHYMOTRYPSIN-EGLIN C COMPLEX AT 2.0 ANGSTROMS RESOLUTION
1MEE 1992-10-15 2.0 THE COMPLEX BETWEEN THE SUBTILISIN FROM A MESOPHILIC BACTERIUM AND THE LEECH INHIBITOR EGLIN-C
1EGP 1995-12-07 2.0 PROTEINASE INHIBITOR EGLIN C WITH HYDROLYSED REACTIVE CENTER
1P2O 2004-04-20 2.0 Structural consequences of accommodation of four non-cognate amino-acid residues in the S1 pocket of bovine trypsin and chymotrypsin
1N8O 2002-12-18 2.0 Crystal structure of a complex between bovine chymotrypsin and ecotin
1GL1 2001-11-28 2.1 structure of the complex between bovine alpha-chymotrypsin and PMP-C, an inhibitor from the insect Locusta migratoria
1SBN 1994-01-31 2.1 REFINED CRYSTAL STRUCTURES OF SUBTILISIN NOVO IN COMPLEX WITH WILD-TYPE AND TWO MUTANT EGLINS. COMPARISON WITH OTHER SERINE PROTEINASE INHIBITOR COMPLEXES
6GCH 1990-10-15 2.1 STRUCTURE OF CHYMOTRYPSIN-*TRIFLUOROMETHYL KETONE INHIBITOR COMPLEXES. COMPARISON OF SLOWLY AND RAPIDLY EQUILIBRATING INHIBITORS
1CA0 1997-07-23 2.1 BOVINE CHYMOTRYPSIN COMPLEXED TO APPI
4VGC 1997-11-12 2.1 GAMMA-CHYMOTRYPSIN D-NAPHTHYL-1-ACETAMIDO BORONIC ACID INHIBITOR COMPLEX
1GMH 1994-09-30 2.1 REFINED CRYSTAL STRUCTURE OF 'AGED' AND 'NON-AGED' ORGANOPHOSPHORYL CONJUGATES OF GAMMA-CHYMOTRYPSIN
5J4S 2017-04-12 2.1 alpha-chymotrypsin from bovine pancreas in complex with a modified Bowman-Birk inhibitor from soybean
1DLK 2000-05-03 2.14 CRYSTAL STRUCTURE ANALYSIS OF DELTA-CHYMOTRYPSIN BOUND TO A PEPTIDYL CHLOROMETHYL KETONE INHIBITOR
4Q2K 2014-07-23 2.2 Bovine alpha chymotrypsin bound to a cyclic peptide inhibitor, 5b
1OXG 2004-05-18 2.2 Crystal structure of a complex formed between organic solvent treated bovine alpha-chymotrypsin and its autocatalytically produced highly potent 14-residue peptide at 2.2 resolution
1GMC 1993-10-31 2.2 THE X-RAY CRYSTAL STRUCTURE OF THE TETRAHEDRAL INTERMEDIATE OF GAMMA-CHYMOTRYPSIN IN HEXANE
1GMD 1993-10-31 2.2 X-ray crystal structure of gamma-chymotrypsin in hexane
1TEC 1989-10-15 2.2 CRYSTALLOGRAPHIC REFINEMENT BY INCORPORATION OF MOLECULAR DYNAMICS. THE THERMOSTABLE SERINE PROTEASE THERMITASE COMPLEXED WITH EGLIN-C
1GHA 1994-06-22 2.2 A SECOND ACTIVE SITE IN CHYMOTRYPSIN? THE X-RAY CRYSTAL STRUCTURE OF N-ACETYL-D-TRYPTOPHAN BOUND TO GAMMA-CHYMOTRYPSIN
1CGI 1993-10-31 2.3 THREE-DIMENSIONAL STRUCTURE OF THE COMPLEXES BETWEEN BOVINE CHYMOTRYPSINOGEN*A AND TWO RECOMBINANT VARIANTS OF HUMAN PANCREATIC SECRETORY TRYPSIN INHIBITOR (KAZAL-TYPE)
1HJA 1998-01-14 2.3 LYS 18 VARIANT OF TURKEY OVOMUCOID INHIBITOR THIRD DOMAIN COMPLEXED WITH ALPHA-CHYMOTRYPSIN
1CGJ 1993-10-31 2.3 THREE-DIMENSIONAL STRUCTURE OF THE COMPLEXES BETWEEN BOVINE CHYMOTRYPSINOGEN*A AND TWO RECOMBINANT VARIANTS OF HUMAN PANCREATIC SECRETORY TRYPSIN INHIBITOR (KAZAL-TYPE)
5J4Q 2017-04-12 2.3 alpha-chymotrypsin from bovine pancreas in complex with Bowman-Birk inhibitor from soybean
1SIB 1993-10-31 2.4 REFINED CRYSTAL STRUCTURES OF SUBTILISIN NOVO IN COMPLEX WITH WILD-TYPE AND TWO MUTANT EGLINS. COMPARISON WITH OTHER SERINE PROTEINASE INHIBITOR COMPLEXES
3BG4 2008-07-29 2.5 The crystal structure of guamerin in complex with chymotrypsin and the development of an elastase-specific inhibitor
1CHG 1976-11-22 2.5 CHYMOTRYPSINOGEN,2.5 ANGSTROMS CRYSTAL STRUCTURE, COMPARISON WITH ALPHA-CHYMOTRYPSIN,AND IMPLICATIONS FOR ZYMOGEN ACTIVATION
1CBW 1997-07-23 2.6 BOVINE CHYMOTRYPSIN COMPLEXED TO BPTI
5GCH 1990-10-15 2.7 CHEMISTRY OF CAGED ENZYMES /II$. PHOTOACTIVATION OF INHIBITED CHYMOTRYPSIN
2Y6T 2011-04-20 2.74 Molecular Recognition of Chymotrypsin by the Serine Protease Inhibitor Ecotin from Yersinia pestis
1MTN 1996-08-17 2.8 BOVINE ALPHA-CHYMOTRYPSIN:BPTI CRYSTALLIZATION
1EX3 2000-05-17 3.0 CRYSTAL STRUCTURE OF BOVINE CHYMOTRYPSINOGEN A (TETRAGONAL)
1GL0 2001-11-28 3.0 structure of the complex between bovine alpha-chymotrypsin and PMP-D2v, an inhibitor from the insect Locusta migratoria

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 E Chymotrypsinogen A P00766 CTRA_BOVIN
100.0 I Chymotrypsinogen A P01051 ICIC_HIRME