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
1YAL 1996-06-20T00:00:00+0000 1.7 CARICA PAPAYA CHYMOPAPAIN AT 1.7 ANGSTROMS RESOLUTION
1BP4 1998-08-12T00:00:00+0000 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.
1BQI 1998-08-16T00:00:00+0000 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.
1CVZ 1999-08-24T00:00:00+0000 1.7 CRYSTAL STRUCTURE ANALYSIS OF PAPAIN WITH CLIK148(CATHEPSIN L SPECIFIC INHIBITOR)
1KHP 2001-11-30T00:00:00+0000 2.0 Monoclinic form of papain/ZLFG-DAM covalent complex
1KHQ 2001-11-30T00:00:00+0000 1.6 ORTHORHOMBIC FORM OF PAPAIN/ZLFG-DAM COVALENT COMPLEX
1PAD 1976-11-01T00:00:00+0000 2.8 Binding of chloromethyl ketone substrate analogues to crystalline papain
1PE6 1991-05-14T00:00:00+0000 2.1 REFINED X-RAY STRUCTURE OF PAPAIN(DOT)E-64-C COMPLEX AT 2.1-ANGSTROMS RESOLUTION
1PIP 1992-10-03T00:00:00+0000 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
1POP 1993-06-24T00:00:00+0000 2.1 X-RAY CRYSTALLOGRAPHIC STRUCTURE OF A PAPAIN-LEUPEPTIN COMPLEX

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