PDBsum entry 1s3d

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Oxidoreductase PDB id
Protein chain
138 a.a. *
SO4 ×4
_CS ×5
Waters ×335
* Residue conservation analysis
PDB id:
Name: Oxidoreductase
Title: Arsenate reductase r60a mutant from e. Coli
Structure: Arsenate reductase. Chain: a. Fragment: r60a. Synonym: arsenical pump modifier. Ec:
Source: Escherichia coli. Organism_taxid: 562. Plasmid: r773
1.54Å     R-factor:   0.138     R-free:   0.199
Authors: S.Demel,B.F.Edwards
Key ref:
S.DeMel et al. (2004). Arginine 60 in the ArsC arsenate reductase of E. coli plasmid R773 determines the chemical nature of the bound As(III) product. Protein Sci, 13, 2330-2340. PubMed id: 15295115 DOI: 10.1110/ps.04787204
13-Jan-04     Release date:   15-Feb-05    
Go to PROCHECK summary

Protein chain
Pfam   ArchSchema ?
P08692  (ARSC1_ECOLX) -  Arsenate reductase
141 a.a.
138 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Enzyme reactions 
   Enzyme class: E.C.  - Arsenate reductase (glutaredoxin).
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Arsenate + glutaredoxin = arsenite + glutaredoxin disulfide + H2O
+ glutaredoxin
= arsenite
+ glutaredoxin disulfide
+ H(2)O
      Cofactor: Mo cation
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     oxidation-reduction process   2 terms 
  Biochemical function     oxidoreductase activity     2 terms  


    Added reference    
DOI no: 10.1110/ps.04787204 Protein Sci 13:2330-2340 (2004)
PubMed id: 15295115  
Arginine 60 in the ArsC arsenate reductase of E. coli plasmid R773 determines the chemical nature of the bound As(III) product.
S.DeMel, J.Shi, P.Martin, B.P.Rosen, B.F.Edwards.
Arsenic is a ubiquitous environmental toxic metal. Consequently, organisms detoxify arsenate by reduction to arsenite, which is then excreted or sequestered. The ArsC arsenate reductase from Escherichia coli plasmid R773, the best characterized arsenic-modifying enzyme, has a catalytic cysteine, Cys 12, in the active site, surrounded by an arginine triad composed of Arg 60, Arg 94, and Arg 107. During the reaction cycle, the native enzyme forms a unique monohydroxyl Cys 12-thiol-arsenite adduct that contains a positive charge on the arsenic. We hypothesized previously that this unstable intermediate allows for rapid dissociation of the product arsenite. In this study, the role of Arg 60 in product formation was evaluated by mutagenesis. A total of eight new structures of ArsC were determined at resolutions between 1.3 A and 1.8 A, with R(free) values between 0.18 and 0.25. The crystal structures of R60K and R60A ArsC equilibrated with the product arsenite revealed a covalently bound Cys 12-thiol-dihydroxyarsenite without a charge on the arsenic atom. We propose that this intermediate is more stable than the monohydroxyarsenite intermediate of the native enzyme, resulting in slow release of product and, consequently, loss of activity.
  Selected figure(s)  
Figure 4.
Figure 4. Interactions of the Csr12 arsonocysteine adducts. Csr12 and its adjacent residues within 4.0 are shown for (A) native ArsC and (B) the R60K mutant. The hydrogen bonds are depicted with dotted lines. The cyan atom in Csr12 is arsenic.
Figure 5.
Figure 5. Reaction mechanism of the R733 ArsC arsenate reductase. The mechanism is consistent with the crystal structures described in Table 1 Go-. In step 1, the free enzyme (structure I) forms the observed covalent intermediate with arsenate (Martin et al. 2001). In step 2, this intermediate is glutathionylated, a structure that has not yet been obtained. In step 3, As(V) is reduced to As(III), producing a dihydroxy arsenite intermediate (structures VI, IX). In step 4, the novel monohydroxy intermediate with a positively charged arsenic is formed (Martin et al. 2001). Finally, in step 5, the free enzyme is regenerated (structure I).
  The above figures are reprinted by permission from the Protein Society: Protein Sci (2004, 13, 2330-2340) copyright 2004.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
19474219 B.Németi, and Z.Gregus (2009).
Mechanism of thiol-supported arsenate reduction mediated by phosphorolytic-arsenolytic enzymes: I. The role of arsenolysis.
  Toxicol Sci, 110, 270-281.  
19286650 E.Ordóñez, K.Van Belle, G.Roos, S.De Galan, M.Letek, J.A.Gil, L.Wyns, L.M.Mateos, and J.Messens (2009).
Arsenate reductase, mycothiol, and mycoredoxin concert thiol/disulfide exchange.
  J Biol Chem, 284, 15107-15116.  
19587982 T.T.Ngu, and M.J.Stillman (2009).
Metal-binding mechanisms in metallothioneins.
  Dalton Trans, (), 5425-5433.  
16607668 G.Roos, S.Loverix, E.Brosens, K.Van Belle, L.Wyns, P.Geerlings, and J.Messens (2006).
The activation of electrophile, nucleophile and leaving group during the reaction catalysed by pI258 arsenate reductase.
  Chembiochem, 7, 981-989.  
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.