PDBsum entry 1eog

Transferase

PDB id: 1eog

Contents

Protein chain

208 a.a. *

Waters ×117

* Residue conservation analysis

PDB id:

1eog

Name:

Transferase

Title:

Crystal structure of pi class glutathione transferase

Structure:

Glutathione s-transferase. Chain: a, b. Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 562. Expression_system_organ: liver.

Biol. unit:

Dimer (from PQS)

Resolution:

2.10Å R-factor: 0.204 R-free: 0.238

Authors:

J.Rossjohn,W.J.Mckinstry,A.J.Oakley,M.W.Parker,G.Stenberg, B.Mannervik,B.Dragani,R.Cocco,A.Aceto

Key ref:

J.Rossjohn et al. (2000). Structures of thermolabile mutants of human glutathione transferase P1-1. J Mol Biol, 302, 295-302. PubMed id: 10970734 DOI: 10.1006/jmbi.2000.4054

Date:

22-Mar-00 Release date: 18-Oct-00

Protein chains

P09211  (GSTP1_HUMAN) -  Glutathione S-transferase P from Homo sapiens

Seq: 210 a.a.

Struc: 208 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.2.5.1.18 - glutathione transferase.
• Reaction: RX + glutathione = an S-substituted glutathione + a halide anion + H⁺

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Added reference

DOI no: 10.1006/jmbi.2000.4054

J Mol Biol 302:295-302 (2000)

PubMed id: 10970734

Structures of thermolabile mutants of human glutathione transferase P1-1.

J.Rossjohn, W.J.McKinstry, A.J.Oakley, M.W.Parker, G.Stenberg, B.Mannervik, B.Dragani, R.Cocco, A.Aceto.

ABSTRACT

An N-capping box motif (Ser/Thr-Xaa-Xaa-Asp) is strictly conserved at the beginning of helix alpha6 in the core of virtually all glutathione transferases (GST) and GST-related proteins. It has been demonstrated that this local motif is important in determining the alpha-helical propensity of the isolated alpha6-peptide and plays a crucial role in the folding and stability of GSTs. Its removal by site-directed mutagenesis generated temperature-sensitive folding mutants unable to refold at physiological temperature (37 degrees C). In the present work, variants of human GSTP1-1 (S150A and D153A), in which the capping residues have been substituted by alanine, have been generated and purified for structural analysis. Thus, for the first time, temperature-sensitive folding mutants of an enzyme, expressed at a permissive temperature, have been crystallized and their three-dimensional structures determined by X-ray crystallography. The crystal structures of human pi class GST temperature-sensitive mutants provide a basis for understanding the structural origin of the dramatic effects observed on the overall stability of the enzyme at higher temperatures upon single substitution of a capping residue.

Selected figure(s)

Figure 1.
Figure 1. Ribbon picture of a monomer of human pi class GST. The location of helix a6, the GST motif II (the conserved sequence motif consisting of helix a6 and the preceding long loop), GSH and the sites of mutation (shown in ball-and-stick) are indicated. This Figure was produced using MOLSCRIPT [Kraulis 1991].

Figure 2.
Figure 2. Stereoviews of the region in GST P1-1 about the sites of mutation. The N-terminal end of helix a6 is shown in ribbon representation and key residues are shown as ball-and-stick. (a) Wild-type structure (9GSS; [Oakley et al 1997]), (b) S150A and (c) D153A. The Figure was produced using MOLSCRIPT [Kraulis 1991].

The above figures are reprinted by permission from Elsevier: J Mol Biol (2000, 302, 295-302) copyright 2000.