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
1YPH 2006-02-14 1.34 High resolution structure of bovine alpha-chymotrypsin
1GG6 2000-09-20 1.4 CRYSTAL STRUCTURE OF GAMMA CHYMOTRYPSIN WITH N-ACETYL-PHENYLALANINE TRIFLUOROMETHYL KETONE BOUND AT THE ACTIVE SITE
1GGD 2000-09-20 1.5 CRYSTAL STRUCTURE OF GAMMA CHYMOTRYPSIN WITH N-ACETYL-LEUCIL-PHENYLALANINE ALDEHYDE BOUND AT THE ACTIVE SITE
2P8O 2007-05-08 1.5 Crystal Structure of a Benzohydroxamic Acid/Vanadate complex bound to chymotrypsin A
3GCT 1991-10-15 1.6 STRUCTURE OF GAMMA-*CHYMOTRYPSIN IN THE RANGE $P*H 2.0 TO $P*H 10.5 SUGGESTS THAT GAMMA-CHYMOTRYPSIN IS A COVALENT ACYL-ENZYME ADDUCT AT LOW $P*H
1GCT 1991-10-15 1.6 IS GAMMA-CHYMOTRYPSIN A TETRAPEPTIDE ACYL-ENZYME ADDUCT OF GAMMA-CHYMOTRYPSIN?
8GCH 1992-04-15 1.6 GAMMA-CHYMOTRYPSIN IS A COMPLEX OF ALPHA-CHYMOTRYPSIN WITH ITS OWN AUTOLYSIS PRODUCTS
1AB9 1997-08-20 1.6 CRYSTAL STRUCTURE OF BOVINE GAMMA-CHYMOTRYPSIN
5CHA 1985-04-01 1.67 THE REFINEMENT AND THE STRUCTURE OF THE DIMER OF ALPHA-*CHYMOTRYPSIN AT 1.67-*ANGSTROMS RESOLUTION
3VGC 1997-11-12 1.67 GAMMA-CHYMOTRYPSIN L-NAPHTHYL-1-ACETAMIDO BORONIC ACID ACID INHIBITOR COMPLEX
4CHA 1985-04-01 1.68 STRUCTURE OF ALPHA-*CHYMOTRYPSIN REFINED AT 1.68 ANGSTROMS RESOLUTION
3RU4 2012-08-29 1.68 Crystal structure of the Bowman-Birk serine protease inhibitor BTCI in complex with trypsin and chymotrypsin
1T8O 2005-03-08 1.7 CRYSTAL STRUCTURE OF THE P1 TRP BPTI MUTANT- BOVINE CHYMOTRYPSIN COMPLEX
1T8L 2005-03-08 1.75 CRYSTAL STRUCTURE OF THE P1 MET BPTI MUTANT- BOVINE CHYMOTRYPSIN COMPLEX
1P2M 2004-04-20 1.75 Structural consequences of accommodation of four non-cognate amino-acid residues in the S1 pocket of bovine trypsin and chymotrypsin
1T8N 2005-03-08 1.75 CRYSTAL STRUCTURE OF THE P1 THR BPTI MUTANT- BOVINE CHYMOTRYPSIN COMPLEX
1P2N 2004-04-20 1.8 Structural consequences of accommodation of four non-cognate amino-acid residues in the S1 pocket of bovine trypsin and chymotrypsin
2VGC 1997-11-12 1.8 GAMMA-CHYMOTRYPSIN D-PARA-CHLORO-1-ACETAMIDO BORONIC ACID INHIBITOR COMPLEX
2GMT 1994-11-01 1.8 THREE-DIMENSIONAL STRUCTURE OF CHYMOTRYPSIN INACTIVATED WITH (2S) N-ACETYL-L-ALANYL-L-PHENYLALANYL-CHLOROETHYL KETONE: IMPLICATIONS FOR THE MECHANISM OF INACTIVATION OF SERINE PROTEASES BY CHLOROKETONES
2CGA 1987-04-16 1.8 BOVINE CHYMOTRYPSINOGEN A. X-RAY CRYSTAL STRUCTURE ANALYSIS AND REFINEMENT OF A NEW CRYSTAL FORM AT 1.8 ANGSTROMS RESOLUTION
1CHO 1988-07-16 1.8 CRYSTAL AND MOLECULAR STRUCTURES OF THE COMPLEX OF ALPHA-*CHYMOTRYPSIN WITH ITS INHIBITOR TURKEY OVOMUCOID THIRD DOMAIN AT 1.8 ANGSTROMS RESOLUTION
2GCT 1991-10-15 1.8 STRUCTURE OF GAMMA-CHYMOTRYPSIN IN THE RANGE PH 2.0 TO PH 10.5 SUGGESTS THAT GAMMA-CHYMOTRYPSIN IS A COVALENT ACYL-ENZYME ADDUCT AT LOW PH
1T8M 2005-03-08 1.8 CRYSTAL STRUCTURE OF THE P1 HIS BPTI MUTANT- BOVINE CHYMOTRYPSIN COMPLEX
1AFQ 1997-09-17 1.8 CRYSTAL STRUCTURE OF BOVINE GAMMA-CHYMOTRYPSIN COMPLEXED WITH A SYNTHETIC INHIBITOR
7GCH 1990-10-15 1.8 STRUCTURE OF CHYMOTRYPSIN-*TRIFLUOROMETHYL KETONE INHIBITOR COMPLEXES. COMPARISON OF SLOWLY AND RAPIDLY EQUILIBRATING INHIBITORS
1K2I 2001-12-05 1.8 Crystal Structure of Gamma-Chymotrypsin in Complex with 7-Hydroxycoumarin
6CHA 1987-04-16 1.8 STRUCTURE OF A TETRAHEDRAL TRANSITION STATE COMPLEX OF ALPHA-*CHYMOTRYPSIN AT 1.8-*ANGSTROMS RESOLUTION
1P2Q 2004-04-20 1.8 Structural consequences of accommodation of four non-cognate amino-acid residues in the S1 pocket of bovine trypsin and chymotrypsin
1T7C 2005-03-08 1.85 CRYSTAL STRUCTURE OF THE P1 GLU BPTI MUTANT- BOVINE CHYMOTRYPSIN COMPLEX
6DI8 2019-01-23 1.86 Crystal structure of bovine alpha-chymotrypsin in space group P65
4GCH 1990-10-15 1.9 STRUCTURE AND ACTIVITY OF TWO PHOTOREVERSIBLE CINNAMATES BOUND TO CHYMOTRYPSIN
1GCD 1994-06-22 1.9 REFINED CRYSTAL STRUCTURE OF 'AGED' AND 'NON-AGED' ORGANOPHOSPHORYL CONJUGATES OF GAMMA-CHYMOTRYPSIN
3GCH 1990-10-15 1.9 CHEMISTRY OF CAGED ENZYMES. BINDING OF PHOTOREVERSIBLE CINNAMATES TO CHYMOTRYPSIN
2GCH 1980-07-09 1.9 REFINED CRYSTAL STRUCTURE OF GAMMA-CHYMOTRYPSIN AT 1.9 ANGSTROMS RESOLUTION
1VGC 1997-11-12 1.9 GAMMA-CHYMOTRYPSIN L-PARA-CHLORO-1-ACETAMIDO BORONIC ACID INHIBITOR COMPLEX
3T62 2012-08-01 2.0 Crystal structure of recombinant Kunitz Type serine protease Inhibitor-1 from the Caribbean Sea anemone Stichodactyla helianthus in complex with bovine chymotrypsin
1N8O 2002-12-18 2.0 Crystal structure of a complex between bovine chymotrypsin and ecotin
1ACB 1993-10-31 2.0 CRYSTAL AND MOLECULAR STRUCTURE OF THE BOVINE ALPHA-CHYMOTRYPSIN-EGLIN C COMPLEX AT 2.0 ANGSTROMS RESOLUTION
2CHA 1977-01-18 2.0 THE STRUCTURE OF CRYSTALLINE ALPHA-CHYMOTRYPSIN, $V.THE ATOMIC STRUCTURE OF TOSYL-ALPHA-CHYMOTRYPSIN AT 2 ANGSTROMS RESOLUTION
1P2O 2004-04-20 2.0 Structural consequences of accommodation of four non-cognate amino-acid residues in the S1 pocket of bovine trypsin and chymotrypsin
1GHB 1994-06-22 2.0 A SECOND ACTIVE SITE IN CHYMOTRYPSIN? THE X-RAY CRYSTAL STRUCTURE OF N-ACETYL-D-TRYPTOPHAN BOUND TO GAMMA-CHYMOTRYPSIN
1GL1 2001-11-28 2.1 structure of the complex between bovine alpha-chymotrypsin and PMP-C, an inhibitor from the insect Locusta migratoria
5J4S 2017-04-12 2.1 alpha-chymotrypsin from bovine pancreas in complex with a modified Bowman-Birk inhibitor from soybean
6GCH 1990-10-15 2.1 STRUCTURE OF CHYMOTRYPSIN-*TRIFLUOROMETHYL KETONE INHIBITOR COMPLEXES. COMPARISON OF SLOWLY AND RAPIDLY EQUILIBRATING INHIBITORS
1CA0 1997-07-23 2.1 BOVINE CHYMOTRYPSIN COMPLEXED TO APPI
4VGC 1997-11-12 2.1 GAMMA-CHYMOTRYPSIN D-NAPHTHYL-1-ACETAMIDO BORONIC ACID INHIBITOR COMPLEX
1GMH 1994-09-30 2.1 REFINED CRYSTAL STRUCTURE OF 'AGED' AND 'NON-AGED' ORGANOPHOSPHORYL CONJUGATES OF GAMMA-CHYMOTRYPSIN
1DLK 2000-05-03 2.14 CRYSTAL STRUCTURE ANALYSIS OF DELTA-CHYMOTRYPSIN BOUND TO A PEPTIDYL CHLOROMETHYL KETONE INHIBITOR
1GHA 1994-06-22 2.2 A SECOND ACTIVE SITE IN CHYMOTRYPSIN? THE X-RAY CRYSTAL STRUCTURE OF N-ACETYL-D-TRYPTOPHAN BOUND TO GAMMA-CHYMOTRYPSIN
1GMC 1993-10-31 2.2 THE X-RAY CRYSTAL STRUCTURE OF THE TETRAHEDRAL INTERMEDIATE OF GAMMA-CHYMOTRYPSIN IN HEXANE
4Q2K 2014-07-23 2.2 Bovine alpha chymotrypsin bound to a cyclic peptide inhibitor, 5b
1GMD 1993-10-31 2.2 X-ray crystal structure of gamma-chymotrypsin in hexane
1OXG 2004-05-18 2.2 Crystal structure of a complex formed between organic solvent treated bovine alpha-chymotrypsin and its autocatalytically produced highly potent 14-residue peptide at 2.2 resolution
1HJA 1998-01-14 2.3 LYS 18 VARIANT OF TURKEY OVOMUCOID INHIBITOR THIRD DOMAIN COMPLEXED WITH ALPHA-CHYMOTRYPSIN
1CGI 1993-10-31 2.3 THREE-DIMENSIONAL STRUCTURE OF THE COMPLEXES BETWEEN BOVINE CHYMOTRYPSINOGEN*A AND TWO RECOMBINANT VARIANTS OF HUMAN PANCREATIC SECRETORY TRYPSIN INHIBITOR (KAZAL-TYPE)
5J4Q 2017-04-12 2.3 alpha-chymotrypsin from bovine pancreas in complex with Bowman-Birk inhibitor from soybean
1CGJ 1993-10-31 2.3 THREE-DIMENSIONAL STRUCTURE OF THE COMPLEXES BETWEEN BOVINE CHYMOTRYPSINOGEN*A AND TWO RECOMBINANT VARIANTS OF HUMAN PANCREATIC SECRETORY TRYPSIN INHIBITOR (KAZAL-TYPE)
1CHG 1976-11-22 2.5 CHYMOTRYPSINOGEN,2.5 ANGSTROMS CRYSTAL STRUCTURE, COMPARISON WITH ALPHA-CHYMOTRYPSIN,AND IMPLICATIONS FOR ZYMOGEN ACTIVATION
3BG4 2008-07-29 2.5 The crystal structure of guamerin in complex with chymotrypsin and the development of an elastase-specific inhibitor
1CBW 1997-07-23 2.6 BOVINE CHYMOTRYPSIN COMPLEXED TO BPTI
5GCH 1990-10-15 2.7 CHEMISTRY OF CAGED ENZYMES /II$. PHOTOACTIVATION OF INHIBITED CHYMOTRYPSIN
2Y6T 2011-04-20 2.74 Molecular Recognition of Chymotrypsin by the Serine Protease Inhibitor Ecotin from Yersinia pestis
1MTN 1996-08-17 2.8 BOVINE ALPHA-CHYMOTRYPSIN:BPTI CRYSTALLIZATION
1JHN 2001-10-10 2.9 Crystal Structure of the Lumenal Domain of Calnexin
1EX3 2000-05-17 3.0 CRYSTAL STRUCTURE OF BOVINE CHYMOTRYPSINOGEN A (TETRAGONAL)
1GL0 2001-11-28 3.0 structure of the complex between bovine alpha-chymotrypsin and PMP-D2v, an inhibitor from the insect Locusta migratoria

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