Role of electrostatic interactions in 2,2,2-trifluoroethanol-induced structural changes and aggregation of alpha-chymotrypsin.


Abstract

It has been recently demonstrated that alpha-chymotrypsin (CT) can be driven toward amyloid aggregation by addition of 2,2,2-trifluoroethanol (TFE), at intermediate concentrations. In the present article, the process of TFE-induced CT aggregation was investigated in more detailed kinetic terms where the effects of medium conditions, such as temperature, presence of kosmotropic and chaotropic salts, pH and chemical modification of lysine residues were examined. Various techniques, including light scattering, fluorescence and circular dichroism spectroscopy, were used to follow and characterize this process. The kinetics of aggregation was found to obey a second-order reaction with respect to protein concentration. The aggregation-prone A-state and aggregation-deficient TFE- or T-state of CT were found to be induced at lower TFE concentrations in the presence of salts. Use of acidic and alkaline conditions and lysine modification also promoted the formation of the T-state. Results presented suggest a role for electrostatic interactions in the aggregation process. Study holds ProTherm entries: 23978, 23979, 23980, 23981, 23982, 23983 Extra Details: scan rate: 0.5 degressC/min Aggregation; alpha-Chymotrypsin; Trifluoroethanol; Amyloid; Electrostatic interaction; Hydrophobic interaction

Submission Details

ID: sDVkvviC4

Submitter: Connie Wang

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

Version: 1

Publication Details
Rezaei-Ghaleh N;Ebrahim-Habibi A;Moosavi-Movahedi AA;Nemat-Gorgani M,Arch. Biochem. Biophys. (2007) Role of electrostatic interactions in 2,2,2-trifluoroethanol-induced structural changes and aggregation of alpha-chymotrypsin. PMID:17141725
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
99.8 Calnexin P24643 CALX_CANLF
96.4 Calnexin Q5R440 CALX_PONAB
96.1 Calnexin P27824 CALX_HUMAN
97.6 Calnexin P35564 CALX_MOUSE
96.4 Calnexin P35565 CALX_RAT