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PDBsum entry 2caq

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Transferase PDB id
2caq
Jmol
Contents
Protein chain
204 a.a. *
Ligands
GSH
PG4
BME
Waters ×197
* Residue conservation analysis
PDB id:
2caq
Name: Transferase
Title: Structure of r21l mutant of sh28gst in complex with gsh
Structure: Glutathione s-transferase 28 kda. Chain: a. Synonym: gst 28, gst class-sigma. Engineered: yes. Mutation: yes
Source: Schistosoma haematobium. Blood fluke. Organism_taxid: 6185. Expressed in: escherichia coli. Expression_system_taxid: 469008.
Biol. unit: Dimer (from PDB file)
Resolution:
2.00Å     R-factor:   0.190     R-free:   0.240
Authors: P.Baiocco,L.J.Gourlay,F.Angelucci,A.Bellelli,A.E.Miele,M.Bru
Key ref:
P.Baiocco et al. (2006). Probing the mechanism of GSH activation in Schistosoma haematobium glutathione-S-transferase by site-directed mutagenesis and X-ray crystallography. J Mol Biol, 360, 678-689. PubMed id: 16777141 DOI: 10.1016/j.jmb.2006.05.040
Date:
22-Dec-05     Release date:   21-Jun-06    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
P30114  (GST28_SCHHA) -  Glutathione S-transferase class-mu 28 kDa isozyme
Seq:
Struc:
211 a.a.
204 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 7 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.2.5.1.18  - Glutathione transferase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: RX + glutathione = HX + R-S-glutathione
RX
+
glutathione
Bound ligand (Het Group name = GSH)
corresponds exactly
= HX
+ R-S-glutathione
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     metabolic process   1 term 
  Biochemical function     transferase activity     2 terms  

 

 
    reference    
 
 
DOI no: 10.1016/j.jmb.2006.05.040 J Mol Biol 360:678-689 (2006)
PubMed id: 16777141  
 
 
Probing the mechanism of GSH activation in Schistosoma haematobium glutathione-S-transferase by site-directed mutagenesis and X-ray crystallography.
P.Baiocco, L.J.Gourlay, F.Angelucci, J.Fontaine, M.Hervé, A.E.Miele, F.Trottein, M.Brunori, A.Bellelli.
 
  ABSTRACT  
 
During turnover, the catalytic tyrosine residue (Tyr10) of the sigma class Schistosoma haematobium wild-type glutathione-S-transferase is expected to switch alternately in and out of the reduced glutathione-binding site (G-site). The Tyrout10 conformer forms a pi-cation interaction with the guanidinium group of Arg21. As in other similar glutathione-S-transferases, the catalytic Tyr has a low pKa of 7.2. In order to investigate the catalytic role of Tyr10, and the structural and functional roles of Arg21, we carried out structural studies on two Arg21 mutants (R21L and R21Q) and a Tyr10 mutant, Y10F. Our crystallographic data for the two Arg21 mutants indicate that only the Tyrout10 conformation is populated, thereby excluding a role of Arg21 in the stabilisation of the out conformation. However, Arg21 was confirmed to be catalytically important and essential for the low pKa of Tyr10. Upon comparison with structural data generated for reduced glutathione-bound and inhibitor-bound wild-type enzymes, it was observed that the orientations of Tyr10 and Arg35 are concerted and that, upon ligand binding, minor rearrangements occur within conserved residues in the active site loop. These rearrangements are coupled to quaternary rigid-body movements at the dimer interface and alterations in the localisation and structural order of the C-terminal domain.
 
  Selected figure(s)  
 
Figure 1.
Figure 1. The movement of conserved Arg35. The two alternative positions of Arg35 were observed in the structures of (a) GTX-bound wtGST showing only the Tyr^in10 conformer and (b) R21L showing the Tyr^out10 conformer. Arg35 is indicated by green sticks, whereas other important residues are indicated by blue sticks. This Figure was generated using ViewerLite 4.2 (Accelrys Inc.).
Figure 2.
Figure 2. Stereo views of the electron density (2F[o]–F[c] at 1σ) showing the two conformers of Tyr10 in the G-site. (a) R21L, showing Tyr^out10 at H-bond distance from Asp33. The new residue, Leu21 is at a distance of 4 Å from Tyr10. Tyr^out10 hinders the movement of the Arg35 side-chain, as displayed in Figure 1(b). (b) GTX-bound wtGST, showing the polar interaction between the hydroxyl group of the Tyr^in10 and the sulphur atom of GTX. Tyr10, Arg21, Asp33 and Arg35 are orientated as shown in Figure 1(a).
 
  The above figures are reprinted by permission from Elsevier: J Mol Biol (2006, 360, 678-689) copyright 2006.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
19823950 J.J.Tang, M.W.Wang, E.Z.Jia, J.J.Yan, Q.M.Wang, J.Zhu, Z.J.Yang, X.Lu, and L.S.Wang (2010).
The common variant in the GSTM1 and GSTT1 genes is related to markers of oxidative stress and inflammation in patients with coronary artery disease: a case-only study.
  Mol Biol Rep, 37, 405-410.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time.