Heat denaturation of pepsinogen in a water-ethanol mixture.

Abstract

The effect of ethanol and pH on thermodynamic parameters and cooperativity of pepsinogen heat denaturation was studied by scanning microcalorimetry. Addition of 20% ethanol decreases the protein denaturation temperature by 10.7 degrees C at pH 6.4 and 15.8 degrees C at pH 8.0. It also decreases the denaturation heat capacity increment from 5.8 to 4.2 kcal/K.mol. The dependences of calorimetric denaturation enthalpy on denaturation temperature both in water and 20% ethanol are linear and intersect at about 95 degrees C. In 20% ethanol the pH shift from 5.9 to 8.0 results in a decreased number of cooperative domains in pepsinogen. This process causes no changes either in the secondary structure or in the local surroundings of aromatic amino acids. It is concluded that ethanol addition does not affect the cooperativity of pepsinogen denaturation substantially until the pH change provokes redistribution of charges in the protein molecule. Study holds ProTherm entries: 9828, 9829, 9830, 9831, 9832, 9833, 9834, 9835, 9836, 9837, 9838, 9839, 9840 Extra Details: pepsinogen; heat denaturation; ethanol; scanning microcalorimetry

Submission Details

ID: eiChr5fS3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:38 p.m.

Version: 1

Publication Details
Makarov AA;Protasevich II;Bazhulina NP;Esipova NG,FEBS Lett. (1995) Heat denaturation of pepsinogen in a water-ethanol mixture. PMID:8001679
Additional Information

Study Summary

Number of data points 26
Proteins Gastricsin ; Pepsin A
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: dHcal pH:8.2 ; Experimental Assay: Tm pH:8.2 ; Experimental Assay: dHcal pH:7.3 ; Experimental Assay: Tm pH:7.3 ; Experimental Assay: dHcal pH:6.8 ; Experimental Assay: Tm pH:6.8 ; Experimental Assay: dHcal pH:5.9 ; Experimental Assay: Tm pH:5.9 ; Experimental Assay: dHcal pH:8.0 ; Experimental Assay: Tm pH:8.0 ; Experimental Assay: dHcal pH:7.7 ; Experimental Assay: Tm pH:7.7 ; Experimental Assay: dHcal pH:7.2 ; Experimental Assay: Tm pH:7.2 ; Experimental Assay: dHcal pH:6.4 ; Experimental Assay: Tm pH:6.4 ; Experimental Assay: dHcal pH:6.0 ; Experimental Assay: Tm pH:6.0
Libraries Mutations for sequence LVKVPLVRKKSLRQNLIKDGKLKDFLKTHKHNPASKYFPEAAALIGDEPLENYLDTEYFGTIGIGTPAQDFTVIFDTGSSNLWVPSVYCSSLACSDHNQFNPDDSSTFEATSQELSITYGTGSMTGILGYDTVQVGGISDTNQIFGLSETEPGSFLYYAPFDGILGLAYPSISASGATPVFDNLWDQGLVSQDLFSVYLSSNDDSGSVVLLGGIDSSYYTGSLNWVPVSVEGYWQITLDSITMDGETIACSGGCQAIVDTGTSLLTGPTSAIANIQSDIGASENSDGEMVISCSSIDSLPDIVFTIDGVQYPLSPSAYILQDDDSCTSGFEGMDVPTSSGELWILGDVFIRQYYTVFDRANNKVGLAPVA

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
3PSG 1993-01-15 1.65 THE HIGH RESOLUTION CRYSTAL STRUCTURE OF PORCINE PEPSINOGEN
2PSG 1992-10-15 1.8 REFINED STRUCTURE OF PORCINE PEPSINOGEN AT 1.8 ANGSTROMS RESOLUTION
4PEP 1990-04-15 1.8 THE MOLECULAR AND CRYSTAL STRUCTURES OF MONOCLINIC PORCINE PEPSIN REFINED AT 1.8 ANGSTROMS RESOLUTION
3PEP 1990-04-15 2.3 REVISED 2.3 ANGSTROMS STRUCTURE OF PORCINE PEPSIN. EVIDENCE FOR A FLEXIBLE SUBDOMAIN
5PEP 1990-07-15 2.34 X-RAY ANALYSES OF ASPARTIC PROTEASES. II. THREE-DIMENSIONAL STRUCTURE OF THE HEXAGONAL CRYSTAL FORM OF PORCINE PEPSIN AT 2.3 ANGSTROMS RESOLUTION
1F34 2001-02-01 2.45 CRYSTAL STRUCTURE OF ASCARIS PEPSIN INHIBITOR-3 BOUND TO PORCINE PEPSIN
1PSA 1994-01-31 2.9 STRUCTURE OF A PEPSIN(SLASH)RENIN INHIBITOR COMPLEX REVEALS A NOVEL CRYSTAL PACKING INDUCED BY MINOR CHEMICAL ALTERATIONS IN THE INHIBITOR
1YX9 2005-05-24 3.0 Effect of Dimethyl Sulphoxide on the crystal structure of Porcine Pepsin

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Gastricsin P30879 PEPC_PIG
99.5 Pepsin A P00791 PEPA_PIG