PDBsum entry: 1lju

Oxidoreductase

PDB-id

1lju


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Waters ×229

* Residue conservation analysis

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PDB id:

1lju

Name:

Oxidoreductase

Title:

X-ray structure of c15a arsenate reductase from pi258 complexed with arsenite

Structure:

Arsenate reductase. Chain: a. Synonym: arsenical pump modifier. Engineered: yes. Mutation: yes. Other_details: oxidised

Source:

Staphylococcus aureus. Organism_taxid: 1280. Gene: arsc. Expressed in: escherichia coli. Expression_system_taxid: 562.

UniProt:

P0A006 (ARSC_STAAU)

Seq:

131 a.a.

Struc:

131 a.a.*

Key:		PfamA domain
		Secondary structure			CATH domain

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

Enzyme class:

E.C.3.1.3.48 [IntEnz] [ExPASy] [KEGG] [BRENDA]

Reaction:

Protein tyrosine phosphate + H₂O = protein tyrosine + phosphate (see diagram below)

Resolution:

1.40Å

R-factor:

0.241

R-free:

0.267

Authors:

I.Zegers,J.C.Martins,R.Willem,L.Wyns,J.Messens

Key ref:

J.Messens et al. (2002). All intermediates of the arsenate reductase mechanism, including an intramolecular dynamic disulfide cascade.. Proc Natl Acad Sci U S A, 99, 8506-8511. [PubMed id: 12072565] [DOI: 10.1073/pnas.132142799]

Date:

22-Apr-02

Release date:

07-Aug-02

Related entries:

1jf8
x-ray structure of reduced c10s, c15a arsenate reductase
from pi258
1ljl
wild type pi258 s. Aureus arsenate reductase
1lk0
disulfide intermediate of c89l arsenate reductase from pi25

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Enzyme reaction for E.C.3.1.3.48

Protein tyrosine phosphate	+	H(2)O	=	protein tyrosine	+	phosphate

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

Key reference

DOI no: 10.1073/pnas.132142799 Proc Natl Acad Sci U S A 99:8506-8511 (2002)

PubMed id: 12072565

All intermediates of the arsenate reductase mechanism, including an intramolecular dynamic disulfide cascade.

J.Messens, J.C.Martins, K.Van Belle, E.Brosens, A.Desmyter, M.De Gieter, J.M.Wieruszeski, R.Willem, L.Wyns, I.Zegers.

ABSTRACT

The mechanism of pI258 arsenate reductase (ArsC) catalyzed arsenate reduction, involving its P-loop structural motif and three redox active cysteines, has been unraveled. All essential intermediates are visualized with x-ray crystallography, and NMR is used to map dynamic regions in a key disulfide intermediate. Steady-state kinetics of ArsC mutants gives a view of the crucial residues for catalysis. ArsC combines a phosphatase-like nucleophilic displacement reaction with a unique intramolecular disulfide bond cascade. Within this cascade, the formation of a disulfide bond triggers a reversible "conformational switch" that transfers the oxidative equivalents to the surface of the protein, while releasing the reduced substrate.

Selected figure(s)

Figure 3.
Fig. 3. (A) Scheme of the reaction mechanism of pI258 ArsC. (1) The nucleophilic attack of the thiol of Cys-10; (2) the formation of a covalent Cys-10-HAsO[ - ]intermediate; (3) the nucleophilic attack of the thiol of Cys-82 with arsenite release; (4) the formation of a Cys-10-Cys-82 intermediate and the nucleophilic attack of the thiol of Cys-89; (5) the formation of a Cys-82-Cys-89 disulfide. (B-F) A stereo view of the 2F[o] F[c] electron density maps contoured at 1.0 placed next to its corresponding reaction step in A. (B) The P-loop (residues 10-17) in the structure of reduced wild-type ArsC with Cys-10 in the center of the image. The P-loop is fully structured, despite the absence of bound oxyanion (2.0 Å). (C) In the structure of C15A ArsC-HAsO[ - ], an arsenic is covalently bound on Cys-10, surrounded by three oxygens in a plane and a water molecule opposite the sulfur of Cys-10 (1.4 Å). (D) Oxidized ArsC C89L with the intermediate Cys-10-Cys-82 disulfide bond (1.6 Å). (E) A view on the flexible looped-out region of oxidized ArsC C89L, where Cys-89 has left the hydrophobic core and is replaced by Leu-92 upon Cys-10-Cys-82 formation. The electron density in this highly flexible region is not so well defined. (F) A view on the surface of oxidized ArsC C10SC15A (6) with the Cys-82-Cys-89 disulfide bond. Figure 4.
Fig. 4. The movement of the "conformational switch" in the flexible segment (residues 80-98) trapped in four different ArsC crystals. Starting with a helix in the reduced ArsC wild type (blue), via oxidized ArsC C89L (Cys-10-Cys-82) in the first (yellow) and in the second (red) molecule in the asymmetric unit to finally looping out to form the C82-C89 disulfide (green). The two arrows indicate the movement of L92 and C89.

Figures were selected by an automated process.

Literature references that cite this PDB file's key reference

PubMed id Reference

19675666 G.Roos, N.Foloppe, K.Van Laer, L.Wyns, L.Nilsson, P.Geerlings, and J.Messens (2009).
How thioredoxin dissociates its mixed disulfide.

PLoS Comput Biol, 5, e1000461.

19304854 L.López-Maury, A.M.Sánchez-Riego, J.C.Reyes, and F.J.Florencio (2009).
The glutathione/glutaredoxin system is essential for arsenate reduction in Synechocystis sp. strain PCC 6803.

J Bacteriol, 191, 3534-3543.

19424433 S.M.Marino, and V.N.Gladyshev (2009).
A structure-based approach for detection of thiol oxidoreductases and their catalytic redox-active cysteine residues.

PLoS Comput Biol, 5, e1000383.

19598234 S.W.Fan, R.A.George, N.L.Haworth, L.L.Feng, J.Y.Liu, and M.A.Wouters (2009).
Conformational changes in redox pairs of protein structures.

Protein Sci, 18, 1745-1765.

17609383 D.S.Touw, C.E.Nordman, J.A.Stuckey, and V.L.Pecoraro (2007).
Identifying important structural characteristics of arsenic resistance proteins by using designed three-stranded coiled coils.

Proc Natl Acad Sci U S A, 104, 11969-11974.

PDB code: 2jgo

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.

16332858 F.S.Islam, R.L.Pederick, A.G.Gault, L.K.Adams, D.A.Polya, J.M.Charnock, and J.R.Lloyd (2005).
Interactions between the Fe(III)-reducing bacterium Geobacter sulfurreducens and arsenate, and capture of the metalloid by biogenic Fe(II).

Appl Environ Microbiol, 71, 8642-8648.

15691908 S.Silver, and L.T.Phung (2005).
Genes and enzymes involved in bacterial oxidation and reduction of inorganic arsenic.

Appl Environ Microbiol, 71, 599-608.

15159594 J.Messens, I.Van Molle, P.Vanhaesebrouck, K.Van Belle, K.Wahni, J.C.Martins, L.Wyns, and R.Loris (2004).
The structure of a triple mutant of pI258 arsenate reductase from Staphylococcus aureus and its 5-thio-2-nitrobenzoic acid adduct.

Acta Crystallogr D Biol Crystallogr, 60, 1180-1184.

PDB codes: 1rxe 1rxi

12618461 L.I.Leichert, C.Scharf, and M.Hecker (2003).
Global characterization of disulfide stress in Bacillus subtilis.

J Bacteriol, 185, 1967-1975.

12949088 L.López-Maury, F.J.Florencio, and J.C.Reyes (2003).
Arsenic sensing and resistance system in the cyanobacterium Synechocystis sp. strain PCC 6803.

J Bacteriol, 185, 5363-5371.

12777806 P.Retailleau, and T.Prangé (2003).
Phasing power at the K absorption edge of organic arsenic.

Acta Crystallogr D Biol Crystallogr, 59, 887-896.

PDB code: 1n4f

14617642 R.Li, J.D.Haile, and P.J.Kennelly (2003).
An arsenate reductase from Synechocystis sp. strain PCC 6803 exhibits a novel combination of catalytic characteristics.

J Bacteriol, 185, 6780-6789.

12829274 S.Silver (2003).
Bacterial silver resistance: molecular biology and uses and misuses of silver compounds.

FEMS Microbiol Rev, 27, 341-353.

12165430 R.Mukhopadhyay, B.P.Rosen, L.T.Phung, and S.Silver (2002).
Microbial arsenic: from geocycles to genes and enzymes.

FEMS Microbiol Rev, 26, 311-325.

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.