Molecular characterization of an anion pump. The arsA gene product is an arsenite(antimonate)-stimulated ATPase.

Rosen, Barry P.; Weigel, Ulrich; Karkaria, Cyrus E.; Gangola, Preeti

doi:10.1016/s0021-9258(18)69034-9

Cited by 137 publications

(17 citation statements)

References 22 publications

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“…The properties of the fluorescent changes were consistent with the properties of activation and catalysis. The process was specific for ATP in that neither ADP nor AMP gave a response, even though ADP binds to protein (Rosen et al, 1988). Mg2+ was also required for fluorescence enhancement with ATP, and the ArsA forms a light-activated adduct with [ -32 ] ATP only in the presence of Mg2+ (Rosen et al, 1988), suggesting that Mg2+ ATP is the compound recognized.…”

Section: Discussionmentioning

confidence: 99%

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Interaction of substrate and effector binding sites in the ArsA ATPase

Zhou¹,

Liu²,

Rosen³

1995

Biochemistry

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The ars operon of plasmid R773 confers resistance to antimonials and arsenicals in Escherichia coli by encoding an ATP-dependent extrusion system for the oxyanions. The catalytic subunit, the ArsA protein, is an ATPase with two nucleotide binding consensus sequences, one in the N-terminal half and one in the C-terminal half of the protein. The ArsA ATPase is allosterically activated by tricoordinate binding of As(3+) or Sb(3+) to three cysteine thiolates. Previous measurements suggested that the intrinsic fluorescence of tryptophans might be useful for examining binding of Mg2+ ATP and antimonite. In the present study an increase in intrinsic tryptophan fluorescence was observed upon addition of Mg2+ ATP. This enhancement was reversed by addition of antimonite. The ArsA protein contains four tryptophan residues: Trp159, Trp253, Trp522, and Trp524. The first two were altered to tyrosine residues by site-directed mutagenesis. Cells expressing both the arsAW159Y and arsAW253Y mutations retained resistance to arsenite, and the purified W159Y and W253Y proteins retained ATPase activity. While the intrinsic tryptophan fluorescence of the W253Y protein responded to addition of Mg2+ ATP, intrinsic tryptophan fluorescence in the purified W159Y protein was no longer enhanced by substrate. These results suggest that Trp159 is conformationally coupled to one or both of the nucleotide binding sites and provides a useful probe for the interaction of effector and substrate binding sites.

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Section: Discussionmentioning

confidence: 99%

“…When the ArsA subunit is present, the complex is an obligatory ATP-coupled system. When the ars A gene is highly expressed, the ArsA protein can be isolated from the cytosol of E. coli as a soluble ATPase (Rosen et al, 1988).…”

mentioning

confidence: 99%

Interaction of substrate and effector binding sites in the ArsA ATPase

Zhou¹,

Liu²,

Rosen³

1995

Biochemistry

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“…Measurements of Static Fluorescence. ArsA protein was purified by the method of Rosen et al (1988), as modified by Hsu and Rosen (1989b). Static fluorescence measurements were performed with an SLM 8000 spectrofluorometer.…”

Section: Methodsmentioning

confidence: 99%

“…Purified ArsA protein exhibits oxyanion-stimulated ATPase activity and can be photo-cross-linked with [ -32 ] ATP (Rosen et al, 1988). Although ATP hydrolysis required the oxyanionic substrate, nucleotide binding was independent of the presence of oxyanion.…”

mentioning

confidence: 99%

Ligand interactions in the ArsA protein, the catalytic component of an anion-translocating adenosinetriphosphatase

Karkaria¹,

Steiner²,

Rosen³

1991

Biochemistry

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The ars operon of the conjugative R-factor R773 produces resistance to arsenicals in cells of Escherichia coli. The operon encodes an oxyanion pump which is composed of a membrane subunit, the 45.5-kDa ArsB protein, and a catalytic subunit, the 63-kDa ArsA protein. Purified ArsA protein is an arsenite(antimonite)-stimulated ATPase. From its amino acid sequence, as deduced from the nucleotide sequence, the ArsA protein has four tryptophanyl residues which could serve as intrinsic fluorescent probes for the study of substrate-induced conformational changes. Both static and dynamic measurements of tryptophan fluorescence were performed with the ArsA protein. Results from static anisotropy measurements indicated differences in molecular motion with addition of ATP, SbO2-, or Mg2+. These results were supported by time decay measurements of fluorescence anisotropy. The results of time decay measurements indicated a shorter correlation time, reflecting localized motion in the vicinity of the probe, and a longer correlation time, which could have arisen from rotation of the major portion of the molecule. The longer correlation time changed with addition of the various effectors, especially MgCl2, suggesting that binding of Mg2+ decreases probe mobility.

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“…Główną podjednostkę pompy ArsAB stanowi należące do nadrodziny MFS (Major Facilitator Superfamily) białko ArsB, które posiada 12 regionów transbłonowych i zlokalizowane jest w błonie komórkowej, gdzie tworzy kanał dla usuwania arsenu i antymonu z komórki, a także jest miejscem przyłączenia do błony homodimeru podjednostki katalitycznej ArsA, zawierającej konserwatywne sekwencje wiązania ATP i wykazującej aktywność ATPazową indukowaną arseninem i antymoninem [67,81,89]. Komórki posiadające funkcjonalny kompleks ArsAB wymagają do usuwania metaloidu energii chemicznej z hydrolizy ATP [25,26].…”

Section: Operony Arsunclassified

Mechanisms of arsenic toxicity and transport in microorganisms

Mucha

Berezowski

Markowska

2017

Postępy Mikrobiologii - Advancements of Microbiology

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Arsenic is an ubiquitous element present in the environment either through geological or anthropogenic activities. Millions of people all over the world are exposed to arsenic mainly via air, drinking water and food sources, which results in higher incidence of cancer. Several mechanisms by which arsenic compounds induce tumorigenesis have been proposed. Arsenic mediates its toxicity by generating oxidative stress, inducing protein misfolding, promoting genotoxicity, hampering DNA repair and disrupting signal transduction. Thus, all organisms have developed multiple pathways for arsenic detoxification. In this article, we review recent advances in the understanding of arsenic toxicity and its transport routes in prokaryotes and eukaryotes, including a dual role of aquaglyceroporins in the uptake and efflux, active transport out of the cell via secondary ion pumps and sequestration of metalloid-thiol conjugates into vacuoles by primary ABC transporters. We believe that such studies are of high importance due to the increasing usage of arsenic-based drugs in the treatment of certain types of cancer and diseases caused by protozoan parasites as well as for the development of bio-and phytoremediation strategies for metalloid-polluted areas. 1. Introduction. 2. The chemical properties and the presence of arsenic in the environment. 3. Pathways for arsenic uptake. 4. Mechanism of trivalent arsenic toxicity. 4.1. Oxidative stress. 4.2. Arsenic binding to proteins. 4.3. Protein aggregation. 5. Pentavalent arsenic toxicity. 6. Cellular detoxification mechanisms of arsenic compounds. 6.1. ars operons. 6.2. ACR genes. 6.3. Removal of arsenic conjugates by the ABC transporters. 6.4. Bi-directional transport of arsenic. 7. Summary

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Molecular characterization of an anion pump. The arsA gene product is an arsenite(antimonate)-stimulated ATPase.

Cited by 137 publications

References 22 publications

Interaction of substrate and effector binding sites in the ArsA ATPase

Interaction of substrate and effector binding sites in the ArsA ATPase

Ligand interactions in the ArsA protein, the catalytic component of an anion-translocating adenosinetriphosphatase

Mechanisms of arsenic toxicity and transport in microorganisms

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