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PDBsum entry 1lbk

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protein ligands Protein-protein interface(s) links
Transferase PDB id
1lbk
Jmol
Contents
Protein chain
208 a.a. *
Ligands
MES ×2
GSH ×2
SO4
Waters ×450
* Residue conservation analysis
PDB id:
1lbk
Name: Transferase
Title: Crystal structure of a recombinant glutathione transferase, created by replacing the last seven residues of each subunit of the human class pi isoenzyme with the additional c-terminal helix of human class alpha isoenzyme
Structure: Glutathione s-transferase class pi chimaera (coda). Chain: a, b. Engineered: yes. Other_details: the protein is a recombinant glutathione transferase created by replacing the last seven residues in each subunit of human glutathione transferase p1-1 with residues 208-213 of human glutathione transferase a1-1.
Source: Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 562.
Biol. unit: Dimer (from PQS)
Resolution:
1.86Å     R-factor:   0.198     R-free:   0.224
Authors: G.K.W.Kong,C.Micaloni,A.P.Mazzetti,M.Nuccetelli,G.Antonini, L.Stella,W.J.Mckinstry,G.Polekhina,J.Rossjohn,G.Federici, G.Ricci,M.W.Parker,M.Lo Bello
Key ref:
C.Micaloni et al. (2003). Engineering a new C-terminal tail in the H-site of human glutathione transferase P1-1: structural and functional consequences. J Mol Biol, 325, 111-122. PubMed id: 12473455 DOI: 10.1016/S0022-2836(02)01178-6
Date:
04-Apr-02     Release date:   17-Apr-02    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chains
Pfam   ArchSchema ?
P09211  (GSTP1_HUMAN) -  Glutathione S-transferase P
Seq:
Struc:
210 a.a.
208 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 6 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     TRAF2-GSTP1 complex   10 terms 
  Biological process     metabolic process   31 terms 
  Biochemical function     S-nitrosoglutathione binding     8 terms  

 

 
DOI no: 10.1016/S0022-2836(02)01178-6 J Mol Biol 325:111-122 (2003)
PubMed id: 12473455  
 
 
Engineering a new C-terminal tail in the H-site of human glutathione transferase P1-1: structural and functional consequences.
C.Micaloni, G.K.Kong, A.P.Mazzetti, M.Nuccetelli, G.Antonini, L.Stella, W.J.McKinstry, G.Polekhina, J.Rossjohn, G.Federici, G.Ricci, M.W.Parker, M.Lo Bello.
 
  ABSTRACT  
 
We have sought the structural basis for the differing substrate specificities of human glutathione transferase P1-1 (class Pi) and human glutathione transferase A1-1 (class Alpha) by adding an extra helix (helix 9), found in the electrophilic substrate-binding site (H-site) of the human class Alpha enzyme, at the C terminus of the human class Pi enzyme. This class Pi-chimera (CODA) was expressed in Escherichia coli, purified and characterized by kinetic and crystallographic approaches. The presence of the newly engineered tail in the H-site of the human Pi enzyme alters its catalytic properties towards those exhibited by the human Alpha enzyme, as assessed using cumene hydroperoxide (diagnostic for class Alpha enzymes) and ethacrynic acid (diagnostic for class Pi) as co-substrates. There is a change of substrate selectivity in the latter case, as the k(cat)/K(m)(EA) value decreases about 70-fold, compared to that of class Pi. With 1-chloro-2,4-dinitrobenzene as co-substrate there is a loss of catalytic activity to about 2% with respect to that of the Pi enzyme. Crystallographic and kinetic studies of the class Pi-chimera provide important clues to explain these altered catalytic properties. The new helix forms many complimentary interactions with the rest of the protein and re-models the original electrophilic substrate-binding site towards one that is more enclosed, albeit flexible. Of particular note are the interactions between Glu205 of the new tail and the catalytic residues, Tyr7 and Tyr108, and the thiol moiety of glutathione (GSH). These interactions may provide an explanation of the more than one unit increase in the pK(a) value of the GSH thiolate and affect both the turnover number and GSH binding, using 1-chloro-2,4-dinitrobenzene as co-substrate. The data presented are consistent with the engineered tail adopting a highly mobile or disordered state in the apo form of the enzyme.
 
  Selected figure(s)  
 
Figure 6.
Figure 6. Comparisons of the CODA crystal structure. Stereo view of the Ca trace of the superimposed CODA (blue trace) with (A) WT enzyme (orange trace) and (B) Alpha class GST A1-1 (brown trace).[13] The superposition on the Alpha class enzyme was based only on the N-terminal domain of each protein. The Figure was generated using BOBSCRIPT. [53]
Figure 8.
Figure 8. Stereo view of the interactions between CODA and GSH. (A) Major GSH conformer; (B) minor GSH conformer. The Figure was generated using BOBSCRIPT.[53]
 
  The above figures are reprinted by permission from Elsevier: J Mol Biol (2003, 325, 111-122) copyright 2003.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
21134126 G.McManus, M.Costa, A.Canals, M.Coll, and T.J.Mantle (2011).
Site-directed mutagenesis of mouse glutathione transferase P1-1 unlocks masked cooperativity, introduces a novel mechanism for 'ping pong' kinetic behaviour, and provides further structural evidence for participation of a water molecule in proton abstraction from glutathione.
  FEBS J, 278, 273-281.
PDB code: 3o76
19538182 I.Axarli, P.Dhavala, A.C.Papageorgiou, and N.E.Labrou (2009).
Crystal structure of Glycine max glutathione transferase in complex with glutathione: investigation of the mechanism operating by the Tau class glutathione transferases.
  Biochem J, 422, 247-256.
PDB code: 3fhs
19780048 I.Quesada-Soriano, L.J.Parker, A.Primavera, J.M.Casas-Solvas, A.Vargas-Berenguel, C.Barón, C.J.Morton, A.Paola Mazzetti, M.Lo Bello, M.W.Parker, and L.García-Fuentes (2009).
Influence of the H-site residue 108 on human glutathione transferase P1-1 ligand binding: Structure-thermodynamic relationships and thermal stability.
  Protein Sci, 18, 2454-2470.
PDB codes: 3hjm 3hjo 3hkr
16988933 M.Kosloff, G.W.Han, S.S.Krishna, R.Schwarzenbacher, M.Fasnacht, M.A.Elsliger, P.Abdubek, S.Agarwalla, E.Ambing, T.Astakhova, H.L.Axelrod, J.M.Canaves, D.Carlton, H.J.Chiu, T.Clayton, M.DiDonato, L.Duan, J.Feuerhelm, C.Grittini, S.K.Grzechnik, J.Hale, E.Hampton, J.Haugen, L.Jaroszewski, K.K.Jin, H.Johnson, H.E.Klock, M.W.Knuth, E.Koesema, A.Kreusch, P.Kuhn, I.Levin, D.McMullan, M.D.Miller, A.T.Morse, K.Moy, E.Nigoghossian, L.Okach, S.Oommachen, R.Page, J.Paulsen, K.Quijano, R.Reyes, C.L.Rife, E.Sims, G.Spraggon, V.Sridhar, R.C.Stevens, H.van den Bedem, J.Velasquez, A.White, G.Wolf, Q.Xu, K.O.Hodgson, J.Wooley, A.M.Deacon, A.Godzik, S.A.Lesley, and I.A.Wilson (2006).
Comparative structural analysis of a novel glutathioneS-transferase (ATU5508) from Agrobacterium tumefaciens at 2.0 A resolution.
  Proteins, 65, 527-537.
PDB code: 2fno
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. Where a reference describes a PDB structure, the PDB code is shown on the right.