Thermal aggregation of alpha-chymotrypsin: role of hydrophobic and electrostatic interactions.


Abstract

We have recently reported that electrostatic interactions may play a critical role in alcohol-induced aggregation of alpha-chymotrypsin (CT). In the present study, we have investigated the heat-induced aggregation of this protein. Thermal aggregation of CT obeyed a characteristic pattern, with a clear lag phase followed by a sharp rise in turbidity. Intrinsic and ANS fluorescence studies, together with fluorescence quenching by acrylamide, suggested that the hydrophobic patches are more exposed in the denatured conformation. Typical chaperone-like proteins, including alpha- and beta-caseins and alpha-crystalline could inhibit thermal aggregation of CT, and their inhibitory effect was nearly pH-independent (within the pH range of 7-9). This was partially counteracted by alpha-, beta- and especially gamma-cyclodextrins, suggesting that hydrophobic interactions may play a major role. Loss of thermal aggregation at extreme acidic and basic conditions, combined with changes in net charge/pH profile of aggregation upon chemical modification of lysine residues are taken to support concomitant involvement of electrostatic interactions. Study holds ProTherm entries: 23977 Extra Details: scan rate: 0.25-1 degreesC/min alpha-Chymotrypsin; Thermal aggregation; Hydrophobic interactions; Chaperone-like proteins; Kosmotropic and chaotropic ions

Submission Details

ID: awEaduYY

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:55 p.m.

Version: 1

Publication Details
Rezaei-Ghaleh N;Ramshini H;Ebrahim-Habibi A;Moosavi-Movahedi AA;Nemat-Gorgani M,Biophys. Chem. (2008) Thermal aggregation of alpha-chymotrypsin: role of hydrophobic and electrostatic interactions. PMID:17964060
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
1JHN 2001-06-28T00:00:00+0000 2.9 Crystal Structure of the Lumenal Domain of Calnexin
1AB9 1997-02-05T00:00:00+0000 1.6 CRYSTAL STRUCTURE OF BOVINE GAMMA-CHYMOTRYPSIN
1AB9 1997-02-05T00:00:00+0000 1.6 CRYSTAL STRUCTURE OF BOVINE GAMMA-CHYMOTRYPSIN
1AB9 1997-02-05T00:00:00+0000 1.6 CRYSTAL STRUCTURE OF BOVINE GAMMA-CHYMOTRYPSIN
1ACB 1991-11-08T00:00:00+0000 2.0 CRYSTAL AND MOLECULAR STRUCTURE OF THE BOVINE ALPHA-CHYMOTRYPSIN-EGLIN C COMPLEX AT 2.0 ANGSTROMS RESOLUTION
1AFQ 1997-03-12T00:00:00+0000 1.8 CRYSTAL STRUCTURE OF BOVINE GAMMA-CHYMOTRYPSIN COMPLEXED WITH A SYNTHETIC INHIBITOR
1AFQ 1997-03-12T00:00:00+0000 1.8 CRYSTAL STRUCTURE OF BOVINE GAMMA-CHYMOTRYPSIN COMPLEXED WITH A SYNTHETIC INHIBITOR
1AFQ 1997-03-12T00:00:00+0000 1.8 CRYSTAL STRUCTURE OF BOVINE GAMMA-CHYMOTRYPSIN COMPLEXED WITH A SYNTHETIC INHIBITOR
1CA0 1997-01-23T00:00:00+0000 2.1 BOVINE CHYMOTRYPSIN COMPLEXED TO APPI
1CA0 1997-01-23T00:00:00+0000 2.1 BOVINE CHYMOTRYPSIN COMPLEXED TO APPI

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Chymotrypsinogen A P00766 CTRA_BOVIN
96.4 Calnexin P35565 CALX_RAT
97.6 Calnexin P35564 CALX_MOUSE
96.1 Calnexin P27824 CALX_HUMAN
96.4 Calnexin Q5R440 CALX_PONAB
99.8 Calnexin P24643 CALX_CANLF