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PDBsum entry 2gh5

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protein ligands Protein-protein interface(s) links
Oxidoreductase PDB id
2gh5
Jmol
Contents
Protein chain
461 a.a. *
Ligands
PO4 ×4
ELI ×2
FAD ×2
GOL ×12
Waters ×703
* Residue conservation analysis
PDB id:
2gh5
Name: Oxidoreductase
Title: Crystal structure of human glutathione reductase complexed w fluoro-analogue of the menadione derivative m5
Structure: Glutathione reductase, mitochondrial. Chain: a, b. Fragment: glutathione reductase. Synonym: gr. Grase. Engineered: yes. Other_details: alkylation of cys 58 by eli is formed by flu see remark 600
Source: Homo sapiens. Human. Organism_taxid: 9606. Gene: gsr, glur, grd1. Expressed in: escherichia coli. Expression_system_taxid: 562
Biol. unit: Dimer (from PQS)
Resolution:
1.70Å     R-factor:   0.224     R-free:   0.256
Authors: K.Fritz-Wolf,A.Winzer,H.Bauer,H.Schirmer,E.Davioud-Charvet
Key ref: H.Bauer et al. (2006). A fluoro analogue of the menadione derivative 6-[2'-(3'-methyl)-1',4'-naphthoquinolyl]hexanoic acid is a suicide substrate of glutathione reductase. Crystal structure of the alkylated human enzyme. J Am Chem Soc, 128, 10784-10794. PubMed id: 16910673 DOI: 10.1021/ja061155v
Date:
25-Mar-06     Release date:   26-Sep-06    
PROCHECK
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 Headers
 References

Protein chains
Pfam   ArchSchema ?
P00390  (GSHR_HUMAN) -  Glutathione reductase, mitochondrial
Seq:
Struc:
 
Seq:
Struc:
522 a.a.
461 a.a.
Key:    PfamA domain  Secondary structure  CATH domain

 Enzyme reactions 
   Enzyme class: E.C.1.8.1.7  - Glutathione-disulfide reductase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: 2 glutathione + NADP+ = glutathione disulfide + NADPH
2 × glutathione
+ NADP(+)
= glutathione disulfide
+ NADPH
      Cofactor: FAD
FAD
Bound ligand (Het Group name = FAD) corresponds exactly
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     cytoplasm   6 terms 
  Biological process     small molecule metabolic process   6 terms 
  Biochemical function     electron carrier activity     6 terms  

 

 
    reference    
 
 
DOI no: 10.1021/ja061155v J Am Chem Soc 128:10784-10794 (2006)
PubMed id: 16910673  
 
 
A fluoro analogue of the menadione derivative 6-[2'-(3'-methyl)-1',4'-naphthoquinolyl]hexanoic acid is a suicide substrate of glutathione reductase. Crystal structure of the alkylated human enzyme.
H.Bauer, K.Fritz-Wolf, A.Winzer, S.Kühner, S.Little, V.Yardley, H.Vezin, B.Palfey, R.H.Schirmer, E.Davioud-Charvet.
 
  ABSTRACT  
 
Glutathione reductase is an important housekeeping enzyme for redox homeostasis both in human cells and in the causative agent of tropical malaria, Plasmodium falciparum. Glutathione reductase inhibitors were shown to have anticancer and antimalarial activity per se and to contribute to the reversal of drug resistance. The development of menadione chemistry has led to the selection of 6-[2'-(3'-methyl)-1',4'-naphthoquinolyl]hexanoic acid, called M(5), as a potent reversible and uncompetitive inhibitor of both human and P. falciparum glutathione reductases. Here we describe the synthesis and kinetic characterization of a fluoromethyl-M(5) analogue that acts as a mechanism-based inhibitor of both enzymes. In the course of enzymatic catalysis, the suicide substrate is activated by one- or two-electron reduction, and then a highly reactive quinone methide is generated upon elimination of the fluorine. Accordingly the human enzyme was found to be irreversibly inactivated with a k(inact) value of 0.4 +/- 0.2 min(-1). The crystal structure of the alkylated enzyme was solved at 1.7 A resolution. It showed the inhibitor to bind covalently to the active site Cys58 and to interact noncovalently with His467', Arg347, Arg37, and Tyr114. On the basis of the crystal structure of the inactivated human enzyme and stopped-flow kinetic studies with two- and four-electron-reduced forms of the unreacted P. falciparum enzyme, a mechanism is proposed which explains naphthoquinone reduction at the flavin of glutathione reductase.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
21554000 B.Tandogan, A.Kuruüzüm-Uz, C.Sengezer, Z.Güvenalp, L...Demirezer, and N.N.Ulusu (2011).
In vitro effects of rosmarinic acid on glutathione reductase and glucose 6-phosphate dehydrogenase.
  Pharm Biol, 49, 587-594.  
21394346 Z.Yan, S.Guang, H.Xu, and X.Liu (2011).
An effective real-time colorimeteric sensor for sensitive and selective detection of cysteine under physiological conditions.
  Analyst, 136, 1916-1921.  
19099057 B.R.Lichtenstein, J.F.Cerda, R.L.Koder, and P.L.Dutton (2009).
Reversible proton coupled electron transfer in a peptide-incorporated naphthoquinone amino acid.
  Chem Commun (Camb), (), 168-170.  
19263098 F.A.de Molfetta, R.F.de Freitas, A.B.da Silva, and C.A.Montanari (2009).
Docking and molecular dynamics simulation of quinone compounds with trypanocidal activity.
  J Mol Model, 15, 1175-1184.  
20161292 S.Espinosa, M.Solivan, and C.P.Vlaar (2009).
Synthesis and redox-enzyme modulation by amino-1,4-dihydro-benzo[d][1,2]dithiine derivatives.
  Tetrahedron Lett, 50, 3023-3026.  
19049979 T.Seefeldt, Y.Zhao, W.Chen, A.S.Raza, L.Carlson, J.Herman, A.Stoebner, S.Hanson, R.Foll, and X.Guan (2009).
Characterization of a novel dithiocarbamate glutathione reductase inhibitor and its use as a tool to modulate intracellular glutathione.
  J Biol Chem, 284, 2729-2737.  
19668867 X.Liu, and S.J.Sturla (2009).
Profiling patterns of glutathione reductase inhibition by the natural product illudin S and its acylfulvene analogues.
  Mol Biosyst, 5, 1013-1024.  
19272349 Y.Zhao, T.Seefeldt, W.Chen, X.Wang, D.Matthees, Y.Hu, and X.Guan (2009).
Effects of glutathione reductase inhibition on cellular thiol redox state and related systems.
  Arch Biochem Biophys, 485, 56-62.  
17579510 A.N.Kuntz, E.Davioud-Charvet, A.A.Sayed, L.L.Califf, J.Dessolin, E.S.Arnér, and D.L.Williams (2007).
Thioredoxin glutathione reductase from Schistosoma mansoni: an essential parasite enzyme and a key drug target.
  PLoS Med, 4, e206.  
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.