Long-range dynamic effects of point mutations propagate through side chains in the serine protease inhibitor eglin c.


Abstract

Long-range interactions are fundamental to protein behaviors such as cooperativity and allostery. In an attempt to understand the role protein flexibility plays in such interactions, the distribution of local fluctuations in a globular protein was monitored in response to localized, nonelectrostatic perturbations. Two valine-to-alanine mutations were introduced into the small serine protease inhibitor eglin c, and the (15)N and (2)H NMR spin relaxation properties of these variants were analyzed in terms of the Lipari-Szabo dynamics formalism and compared to those of the wild type. Significant changes in picosecond to nanosecond dynamics were observed in side chains located as much as 13 A from the point of mutation. Additionally, those residues experiencing altered dynamics appear to form contiguous surfaces within the protein. In the case of V54A, the large-to-small mutation results in a rigidification of connected residues, even though this mutation decreases the global stability. These findings suggest that dynamic perturbations arising from single mutations may propagate away from the perturbed site through networks of interacting side chains. That this is observed in eglin c, a classically nonallosteric protein, suggests that such behavior will be observed in many, if not all, globular proteins. Differences in behavior between the two mutants suggest that dynamic responses will be context-dependent. Study holds ProTherm entries: 18378, 18379, 18380, 18381, 18382, 18383, 18384, 18385 Extra Details: long-range interactions; flexibility; nonelectrostatic perturbations; context-dependent

Submission Details

ID: AUUeRHVq

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:50 p.m.

Version: 1

Publication Details
Clarkson MW;Lee AL,Biochemistry (2004) Long-range dynamic effects of point mutations propagate through side chains in the serine protease inhibitor eglin c. PMID:15449934
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
1EGL 1994-01-31 THE SOLUTION STRUCTURE OF EGLIN C BASED ON MEASUREMENTS OF MANY NOES AND COUPLING CONSTANTS AND ITS COMPARISON WITH X-RAY STRUCTURES
1CSE 1988-07-16 1.2 THE HIGH-RESOLUTION X-RAY CRYSTAL STRUCTURE OF THE COMPLEX FORMED BETWEEN SUBTILISIN CARLSBERG AND EGLIN C, AN ELASTASE INHIBITOR FROM THE LEECH HIRUDO MEDICINALIS. STRUCTURAL ANALYSIS, SUBTILISIN STRUCTURE AND INTERFACE GEOMETRY
1YPH 2006-02-14 1.34 High resolution structure of bovine alpha-chymotrypsin
4B2B 2012-08-01 1.36 Structure of the factor Xa-like trypsin variant triple-Ala (TGPA) in complex with eglin C
1GG6 2000-09-20 1.4 CRYSTAL STRUCTURE OF GAMMA CHYMOTRYPSIN WITH N-ACETYL-PHENYLALANINE TRIFLUOROMETHYL KETONE BOUND AT THE ACTIVE SITE
4B2C 2012-08-01 1.43 Structure of the factor Xa-like trypsin variant triple-Ala (TPA) in complex with eglin C
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
4B1T 2012-08-01 1.78 Structure of the factor Xa-like trypsin variant triple-Ala (TA) in complex with eglin C
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
2SEC 1988-09-07 1.8 STRUCTURAL COMPARISON OF TWO SERINE PROTEINASE-PROTEIN INHIBITOR COMPLEXES. EGLIN-C-SUBTILISIN CARLSBERG AND CI-2-SUBTILISIN NOVO
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
4B2A 2012-08-01 1.89 Structure of the factor Xa-like trypsin variant triple-Ala (TGA) in complex with eglin C
4GCH 1990-10-15 1.9 STRUCTURE AND ACTIVITY OF TWO PHOTOREVERSIBLE CINNAMATES BOUND TO CHYMOTRYPSIN
4H4F 2013-02-27 1.9 Crystal structure of human chymotrypsin C (CTRC) bound to inhibitor eglin c from Hirudo medicinalis
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
2TEC 1992-01-15 1.98 MOLECULAR DYNAMICS REFINEMENT OF A THERMITASE-EGLIN-C COMPLEX AT 1.98 ANGSTROMS RESOLUTION AND COMPARISON OF TWO CRYSTAL FORMS THAT DIFFER IN CALCIUM CONTENT
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
1MEE 1992-10-15 2.0 THE COMPLEX BETWEEN THE SUBTILISIN FROM A MESOPHILIC BACTERIUM AND THE LEECH INHIBITOR EGLIN-C
1ACB 1993-10-31 2.0 CRYSTAL AND MOLECULAR STRUCTURE OF THE BOVINE ALPHA-CHYMOTRYPSIN-EGLIN C COMPLEX AT 2.0 ANGSTROMS RESOLUTION
3TEC 1992-01-15 2.0 CALCIUM BINDING TO THERMITASE. CRYSTALLOGRAPHIC STUDIES OF THERMITASE AT 0, 5 AND 100 MM CALCIUM
1EGP 1995-12-07 2.0 PROTEINASE INHIBITOR EGLIN C WITH HYDROLYSED REACTIVE CENTER
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
6GCH 1990-10-15 2.1 STRUCTURE OF CHYMOTRYPSIN-*TRIFLUOROMETHYL KETONE INHIBITOR COMPLEXES. COMPARISON OF SLOWLY AND RAPIDLY EQUILIBRATING INHIBITORS
5J4S 2017-04-12 2.1 alpha-chymotrypsin from bovine pancreas in complex with a modified Bowman-Birk inhibitor from soybean
1CA0 1997-07-23 2.1 BOVINE CHYMOTRYPSIN COMPLEXED TO APPI
1SBN 1994-01-31 2.1 REFINED CRYSTAL STRUCTURES OF SUBTILISIN NOVO IN COMPLEX WITH WILD-TYPE AND TWO MUTANT EGLINS. COMPARISON WITH OTHER SERINE PROTEINASE INHIBITOR COMPLEXES
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
1TEC 1989-10-15 2.2 CRYSTALLOGRAPHIC REFINEMENT BY INCORPORATION OF MOLECULAR DYNAMICS. THE THERMOSTABLE SERINE PROTEASE THERMITASE COMPLEXED WITH EGLIN-C
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)
1SIB 1993-10-31 2.4 REFINED CRYSTAL STRUCTURES OF SUBTILISIN NOVO IN COMPLEX WITH WILD-TYPE AND TWO MUTANT EGLINS. COMPARISON WITH OTHER SERINE PROTEINASE INHIBITOR COMPLEXES
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
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 E Chymotrypsinogen A P00766 CTRA_BOVIN
100.0 I Chymotrypsinogen A P01051 ICIC_HIRME