Role of the C-terminal helix 9 in the stability and ligandin function of class alpha glutathione transferase A1-1.


Abstract

Helix 9 at the C-terminus of class alpha glutathione transferase (GST) polypeptides is a unique structural feature in the GST superfamily. It plays an important structural role in the catalytic cycle. Its contribution toward protein stability/folding as well as the binding of nonsubstrate ligands was investigated by protein engineering, conformational stability, enzyme activity, and ligand-binding methods. The helix9 sequence displays an unfavorable propensity toward helix formation, but tertiary interactions between the amphipathic helix and the GST seem to contribute sufficient stability to populate the helix on the surface of the protein. The helix's stability is enhanced further by the binding of ligands at the active site. The order of ligand-induced stabilization increases from H-site occupation, to G-site occupation, to the simultaneous occupation of H- and G-sites. Ligand-induced stabilization of helix9 reduces solvent accessible hydrophobic surface by facilitating firmer packing at the hydrophobic interface between helix and GST. This stabilized form exhibits enhanced affinity for the binding of nonsubstrate ligands to ligandin sites (i.e., noncatalytic binding sites). Although helix9 contributes very little toward the global stability of hGSTA1-1, its conformational dynamics have significant implications for the protein's equilibrium unfolding/refolding pathway and unfolding kinetics. Considering the high concentration of reduced glutathione in human cells (about 10 mM), the physiological form of hGSTA1-1 is most likely the thiol-complexed protein with a stabilized helix9. The C-terminus region (including helix9) of the class alpha polypeptide appears not to have been optimized for stability but rather for catalytic and ligandin function. Study holds ProTherm entries: 16309 Extra Details: 1mM EDTA protein stability folding, ligand-binding, C-terminal helix 9,,alpha glutathione transferase A1-1

Submission Details

ID: Z5SDyJuN3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:47 p.m.

Version: 1

Publication Details
Dirr HW;Wallace LA,Biochemistry (1999) Role of the C-terminal helix 9 in the stability and ligandin function of class alpha glutathione transferase A1-1. PMID:10569948
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
6ATR 2018-09-12 1.29 Crystal structure of hGSTA1-1 complexed with two GSH analogues in each subunit
1K3Y 2002-10-30 1.3 Crystal Structure Analysis of human Glutathione S-transferase with S-hexyl glutatione and glycerol at 1.3 Angstrom
1K3L 2002-10-23 1.5 Crystal Structure Analysis of S-hexyl-glutathione Complex of Glutathione Transferase at 1.5 Angstroms Resolution
6ATO 2018-09-12 1.55 Crystal structure of hGSTA1-1 complexed with GSH and MPD in each subunit
5LD0 2016-09-21 1.6 Chimeric GST
6ATP 2018-09-12 1.7 Crystal structure of apo-hGSTA1-1 exhibiting a new conformation of C-terminal helix
1PL1 2004-06-22 1.75 Crystal structure of human glutathione transferase (GST) A1-1 in complex with a decarboxy-glutathione
3KTL 2009-12-22 1.75 Crystal Structure of an I71A human GSTA1-1 mutant in complex with S-hexylglutathione
3Q74 2011-01-12 1.79 Crystal Structure Analysis of the L7A Mutant of the Apo Form of Human Alpha Class Glutathione Transferase
1K3O 2002-10-30 1.8 Crystal Structure Analysis of apo Glutathione S-Transferase
1PL2 2004-06-22 1.8 Crystal structure of human glutathione transferase (GST) A1-1 T68E mutant in complex with decarboxy-glutathione
2R3X 2007-12-18 1.8 Crystal structure of an R15L hGSTA1-1 mutant complexed with S-hexyl-glutathione
2VCV 2008-10-28 1.8 Glutathione transferase A3-3 in complex with glutathione and delta-4- androstene-3-17-dione
1XWG 2005-11-01 1.85 Human GST A1-1 T68E mutant
3ZFB 2012-12-19 1.86 Crystal structure of the I75A mutant of human class alpha glutathione transferase in the apo form
3ZFL 2012-12-19 1.88 Crystal structure of the V58A mutant of human class alpha glutathione transferase in the apo form
5JCU 2016-10-12 1.93 Crystal Structure of hGSTA1-1 with Glutathione Adduct of Phenethyl Isothiocyanate and Cystein Adduct of Phenethyl Isothiocyanate
1YDK 2005-06-07 1.95 Crystal structure of the I219A mutant of human glutathione transferase A1-1 with S-hexylglutathione
3I6A 2009-09-01 1.98 Human GST A1-1 GIMF mutant with Glutathione
1PKW 2004-06-22 2.0 Crystal structure of human glutathione transferase (GST) A1-1 in complex with glutathione
1GSE 1995-09-15 2.0 GLUTATHIONE TRANSFERASE A1-1 COMPLEXED WITH AN ETHACRYNIC ACID GLUTATHIONE CONJUGATE (MUTANT R15K)
6ATQ 2018-09-12 2.0 Crystal structure of apo-hGSTA1-1 exhibiting a new conformation of C-terminal helix
1USB 2004-09-01 2.07 Rational design of a novel enzyme - efficient thioester hydrolysis enabled by the incorporation of a single His residue into human glutathione transferase A1-1
2VCT 2008-10-28 2.1 Glutathione transferase A2-2 in complex with delta-4-andostrene-3-17- dione
4ACS 2011-12-28 2.1 Crystal structure of mutant GST A2-2 with enhanced catalytic efficiency with azathioprine
1PKZ 2004-06-22 2.1 Crystal structure of human glutathione transferase (GST) A1-1
4HJ2 2013-03-27 2.1 Crystal Structure Analysis of GSTA1-1 in complex with chlorambucil
3L0H 2010-01-12 2.13 Crystal Structure Analysis of W21A mutant of human GSTA1-1 in complex with S-hexylglutathione
3IK9 2010-06-23 2.2 Human GST A1-1-GIMF with GSDHN
3U6V 2011-10-26 2.2 Crystal Structure Analysis of L23A mutant of human GST A1-1
2WJU 2009-06-09 2.3 Glutathione transferase A2-2 in complex with glutathione
5LCZ 2016-09-21 2.33 Chimeric GST
3I69 2009-09-01 2.38 Apo Glutathione Transferase A1-1 GIMF-helix mutant
1TDI 2005-01-18 2.4 Crystal Structure of hGSTA3-3 in Complex with Glutathione
1GSD 1995-09-15 2.5 GLUTATHIONE TRANSFERASE A1-1 IN UNLIGANDED FORM
1AGS 1995-07-10 2.5 A SURFACE MUTANT (G82R) OF A HUMAN ALPHA-GLUTATHIONE S-TRANSFERASE SHOWS DECREASED THERMAL STABILITY AND A NEW MODE OF MOLECULAR ASSOCIATION IN THE CRYSTAL
2R6K 2008-08-19 2.51 Crystal structure of an I71V hGSTA1-1 mutant in complex with S-hexylglutathione
1GUH 1993-10-31 2.6 Structure determination and refinement of human alpha class glutathione transferase A1-1, and a comparison with the MU and PI class enzymes
1GSF 1995-09-15 2.7 GLUTATHIONE TRANSFERASE A1-1 COMPLEXED WITH ETHACRYNIC ACID

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.1 Glutathione S-transferase A1 Q7RTV2 GSTA5_HUMAN
90.5 Glutathione S-transferase A1 Q16772 GSTA3_HUMAN
95.0 Glutathione S-transferase A1 P09210 GSTA2_HUMAN
100.0 Glutathione S-transferase A1 P08263 GSTA1_HUMAN