Preparation and crystallization of a<i>Bacillus subtilis</i>arsenate reductase

Guan, Zhi; Hederstedt, Lars; Li, Jinping; Su, Xiao‐Dong

doi:10.1107/s0907444901014020

Cited by 7 publications

(4 citation statements)

References 20 publications

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“…Our Se-Met-substituted crystals diffracted to much higher resolution (14); the Met residues are far away from the active site, and the activity on a PTPase substrate (see below) showed no difference between the wild-type and Se-Met enzymes. Thus, the high-resolution structure of the Se-Met enzyme is reported here.…”

Section: Resultsmentioning

confidence: 96%

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Bacillus subtilis arsenate reductase is structurally and functionally similar to low molecular weight protein tyrosine phosphatases

Bennett

Guan

Laurberg

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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129

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Arsenate is an abundant oxyanion that, because of its ability to mimic the phosphate group, is toxic to cells. Arsenate reductase (EC 1.97.1.5; encoded by the arsC gene in bacteria) participates to achieve arsenate resistance in both prokaryotes and yeast by reducing arsenate to arsenite; the arsenite is then exported by a specific transporter. The crystal structure of Bacillus subtilis arsenate reductase in the reduced form with a bound sulfate ion in its active site is solved at 1.6-Å resolution. Significant structural similarity is seen between arsenate reductase and bovine low molecular weight protein tyrosine phosphatase, despite very low sequence identity. The similarity is especially high between their active sites. It is further confirmed that this structural homology is relevant functionally by showing the phosphatase activity of the arsenate reductase in vitro. Thus, we can understand the arsenate reduction in the light of low molecular weight protein tyrosine phosphatase mechanism and also explain the catalytic roles of essential residues such as Cys-10, Cys-82, Cys-89, Arg-16, and Asp-105. A ''triple cysteine redox relay'' is proposed for the arsenate reduction mechanism.

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Section: Resultsmentioning

confidence: 96%

“…The arsenate reductase (EC 1.97.1.5) is a newly characterized enzyme (11); structural data are accumulating for this diverse family of proteins (12)(13)(14). The B. subtilis arsenate reductase is chosen for crystallographic studies to understand the arsenate resistance mechanism.…”

mentioning

confidence: 99%

Bacillus subtilis arsenate reductase is structurally and functionally similar to low molecular weight protein tyrosine phosphatases

Bennett

Guan

Laurberg

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

129

View full text Add to dashboard Cite

show abstract

“…After increasing the concentration of DTT, the HSQC spectra showed a unique set of peaks corresponding to the pure reduced form of ArsC. Notably, the concentration of DDT used in the crystal preparation was approximately five times that of the enzyme (14,22). In addition, the region from Cys 82 to Val 96 is well defined in the NMR structure of oxidized ArsC.…”

Section: Solution Structures Of the Reduced And Oxidized Forms Of B mentioning

confidence: 90%

Solution Structures and Backbone Dynamics of Arsenate Reductase from Bacillus subtilis

Guo

Peng

et al. 2005

Journal of Biological Chemistry

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Arsenate reductase encoded by the chromosomal arsC gene in Bacillus subtilis catalyzes the intracellular reduction of arsenate to arsenite, which is then extruded from cells through an efficient and specific transport system. Herein, we present the solution structures and backbone dynamics of both the reduced and oxidized forms of arsenate reductase from B. subtilis. The overall structures of both forms are similar to those of bovine low molecular weight protein-tyrosine phosphatase and arsenate reductase from Staphylococcus aureus. However, several features of the tertiary structure and mobility are notably different between the reduced and oxidized forms of B. subtilis arsenate reductase, particularly in the P-loop region and the segment Cys 82 -Cys 89 . The backbone dynamics results demonstrated that the reduced form of arsenate reductase undergoes millisecond conformational changes in the functional P-loop and Cys 82 -Cys 89 , which may facilitate the formation of covalent intermediates and subsequent reduction of arsenate. In the oxidized form, Cys 82 -Cys 89 shows motional flexibility on both picosecond-to-nanosecond and possibly millisecond time scales, which may facilitate the reduction of the oxidized enzyme by thioredoxin to regenerate the active enzyme. Overall, the internal dynamics and static structures of the enzyme provide insights into the molecular mechanism of arsenate reduction, especially the reversible conformational switch and changes in internal motions associated with the catalytic reaction.

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“…However, in view of the coordination environment and the fact that the crystals of Bs _ ArsC were grown in the presence of 100 mM sodium citrate, 23 we may still consider a sodium ion binding site. After changing the rotamer conformation of the side-chain of Asn13 and reconsidering the electron density, a consistent site with five protein oxygen atoms and two well-defined water molecules coordinating a sodium ion is created (Figures 2 and 3(b)).…”

Section: Structure Of Sa _ Arsc H62qmentioning

confidence: 99%

Interplay Between Ion Binding and Catalysis in the Thioredoxin-coupled Arsenate Reductase Family

Roos

Buts

Belle

et al. 2006

Journal of Molecular Biology

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Preparation and crystallization of aBacillus subtilisarsenate reductase

Cited by 7 publications

References 20 publications

Bacillus subtilis arsenate reductase is structurally and functionally similar to low molecular weight protein tyrosine phosphatases

Bacillus subtilis arsenate reductase is structurally and functionally similar to low molecular weight protein tyrosine phosphatases

Solution Structures and Backbone Dynamics of Arsenate Reductase from Bacillus subtilis

Interplay Between Ion Binding and Catalysis in the Thioredoxin-coupled Arsenate Reductase Family

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