Cooperativity in the unfolding transitions of cysteine proteinases. Calorimetric study of the heat denaturation of chymopapain and papain.


Abstract

Differential scanning calorimetry (DSC) was employed to study the thermal unfolding of chymopapain (EC 3.4.22.6) and papain (EC 3.4.22.2), two highly homologous cysteine proteinases from Carica papaya. Under all pH conditions used, both enzymes showed irreversible thermal denaturation. However, results from experiments performed at two different scanning rates suggest that interpretation of data in terms of equilibrium thermodynamics is not unreasonable. For papain, the ratio of calorimetric (delta Hcal) to van't Hoff (delta HvH) enthalpies approximated to 2.0. This value indicates that papain domains unfold almost independently, as it has been reported previously. In contrast, chymopapain displayed a more cooperative behavior with a delta Hcal to delta HvH ratio of 1.3-1.4. DSC curves were analyzed in terms of a mechanism that includes domain-domain interactions. The results showed a negligible interdomain free energy in the case of papain, but a significant value of approx. 1.0 kcal/mol (1 cal = 4.184 J) for chymopapain. These two proteins also differed in the unfolding heat-capacity change, delta Cp, which suggests that their native structures bury different amounts of nonpolar surface area. Study holds ProTherm entries: 9885, 9886, 9887, 9888, 9889, 9890, 9891, 9892, 9893, 9894, 9895, 9896, 9897 Extra Details: cysteine proteinases; papaindomains domain-domaininteractions;,nonpolar surface area

Submission Details

ID: WQeyaUrE3

Submitter: Connie Wang

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

Version: 1

Publication Details
Solís-Mendiola S;Rojo-Domínguez A;Hernández-Arana A,Biochim. Biophys. Acta (1993) Cooperativity in the unfolding transitions of cysteine proteinases. Calorimetric study of the heat denaturation of chymopapain and papain. PMID:8218380
Additional Information

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
2CIO 2006-05-18 1.5 The high resolution x-ray structure of papain complexed with fragments of the Trypanosoma brucei cysteine protease inhibitor ICP.
1PPN 1994-01-31 1.6 STRUCTURE OF MONOCLINIC PAPAIN AT 1.60 ANGSTROMS RESOLUTION
1KHQ 2003-09-09 1.6 ORTHORHOMBIC FORM OF PAPAIN/ZLFG-DAM COVALENT COMPLEX
9PAP 1986-10-24 1.65 STRUCTURE OF PAPAIN REFINED AT 1.65 ANGSTROMS RESOLUTION
1PIP 1993-10-31 1.7 CRYSTAL STRUCTURE OF PAPAIN-SUCCINYL-GLN-VAL-VAL-ALA-ALA-P-NITROANILIDE COMPLEX AT 1.7 ANGSTROMS RESOLUTION: NONCOVALENT BINDING MODE OF A COMMON SEQUENCE OF ENDOGENOUS THIOL PROTEASE INHIBITORS
1YAL 1996-12-23 1.7 CARICA PAPAYA CHYMOPAPAIN AT 1.7 ANGSTROMS RESOLUTION
1CVZ 2000-08-30 1.7 CRYSTAL STRUCTURE ANALYSIS OF PAPAIN WITH CLIK148(CATHEPSIN L SPECIFIC INHIBITOR)
3E1Z 2009-01-27 1.86 Crystal structure of the parasite protesase inhibitor chagasin in complex with papain
1PPP 1994-01-31 1.9 CRYSTAL STRUCTURE OF PAPAIN-E64-C COMPLEX. BINDING DIVERSITY OF E64-C TO PAPAIN S2 AND S3 SUBSITES
4QRV 2015-08-12 1.98 Crystal structure of I86F mutant of papain
1KHP 2003-09-09 2.0 Monoclinic form of papain/ZLFG-DAM covalent complex
1PPD 1985-01-02 2.0 RESTRAINED LEAST-SQUARES REFINEMENT OF THE SULFHYDRYL PROTEASE PAPAIN TO 2.0 ANGSTROMS
3IMA 2010-02-16 2.03 Complex strcuture of tarocystatin and papain
1POP 1993-10-31 2.1 X-RAY CRYSTALLOGRAPHIC STRUCTURE OF A PAPAIN-LEUPEPTIN COMPLEX
6H8T 2018-09-12 2.1 Crystal structure of Papain modify by achiral Ru(II)complex
1PE6 1993-04-15 2.1 REFINED X-RAY STRUCTURE OF PAPAIN(DOT)E-64-C COMPLEX AT 2.1-ANGSTROMS RESOLUTION
1BP4 1999-08-12 2.2 USE OF PAPAIN AS A MODEL FOR THE STRUCTURE-BASED DESIGN OF CATHEPSIN K INHIBITORS. CRYSTAL STRUCTURES OF TWO PAPAIN INHIBITOR COMPLEXES DEMONSTRATE BINDING TO S'-SUBSITES.
1STF 1994-01-31 2.37 THE REFINED 2.4 ANGSTROMS X-RAY CRYSTAL STRUCTURE OF RECOMBINANT HUMAN STEFIN B IN COMPLEX WITH THE CYSTEINE PROTEINASE PAPAIN: A NOVEL TYPE OF PROTEINASE INHIBITOR INTERACTION
1BQI 1999-08-16 2.5 USE OF PAPAIN AS A MODEL FOR THE STRUCTURE-BASED DESIGN OF CATHEPSIN K INHIBITORS. CRYSTAL STRUCTURES OF TWO PAPAIN INHIBITOR COMPLEXES DEMONSTRATE BINDING TO S'-SUBSITES.
4QRG 2015-08-05 2.5 Crystal structure of I86L mutant of papain
3LFY 2010-07-21 2.6 CTD of Tarocystatin in complex with papain
3TNX 2012-09-12 2.62 Structure of the precursor of a thermostable variant of papain at 2.6 Angstroem resolution
5PAD 1977-04-12 2.8 BINDING OF CHLOROMETHYL KETONE SUBSTRATE ANALOGUES TO CRYSTALLINE PAPAIN
2PAD 1977-04-12 2.8 BINDING OF CHLOROMETHYL KETONE SUBSTRATE ANALOGUES TO CRYSTALLINE PAPAIN
1PAD 1977-04-12 2.8 Binding of chloromethyl ketone substrate analogues to crystalline papain
4PAD 1977-04-12 2.8 Binding of chloromethyl ketone substrate analogues to crystalline papain
6PAD 1977-04-12 2.8 Binding of chloromethyl ketone substrate analogues to crystalline papain
4QRX 2015-08-05 3.14 Crystal structure of pro-papain mutant at pH 4.0
3USV 2012-11-28 3.8 Structure of the precursor of a thermostable variant of papain at 3.8 A resolution from a crystal soaked at pH 4

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Papain P00784 PAPA1_CARPA
90.7 Chymopapain P32956 CYSP3_VASCU
100.0 Chymopapain P14080 PAPA2_CARPA