Characterization of the metalloactivation domain of an arsenite/antimonite resistance pump.

Abstract:

:The ArsAB extrusion pump encoded by the ars operon of Escherichia coli plasmid R773 confers resistance to the toxic trivalent metalloids arsenite [As(III)] and antimonite [Sb(III)]. The ArsA ATPase, the catalytic subunit of the pump, has two homologous halves, A1 and A2. At the interface of these two halves are two nucleotide-binding domains and a metalloid-binding domain. Cys-113 and Cys-422 have been shown to form a high-affinity metalloid binding site. The crystal structure of ArsA shows two other bound metalloid atoms, one liganded to Cys-172 and His-453, and the other liganded to His-148 and Ser-420. The contribution of those putative metalloid sites was examined. There was little effect of mutagenesis of residues His-148 and Ser-420 on metalloid binding. However, a C172A ArsA mutant and C172A/H453A double mutant exhibited significantly decreased affinity for Sb(III). These results suggest first that there is only a single high-affinity metalloid binding site in ArsA, and second that Cys-172 controls the affinity of this site for metalloid and hence the efficiency of metalloactivation of the ArsAB efflux pump.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Ruan X,Bhattacharjee H,Rosen BP

doi

10.1111/j.1365-2958.2007.06049.x

subject

Has Abstract

pub_date

2008-01-01 00:00:00

pages

392-402

issue

2

eissn

0950-382X

issn

1365-2958

pii

MMI6049

journal_volume

67

pub_type

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