Molecular characterization of an anion pump. The ArsB protein is the membrane anchor for the ArsA protein.
R-factor mediated bacterial resistance to arsenical salts occurs by active extrusion of the toxic oxyanions from cells of gram negative bacteria. The ars operon of the conjugative plasmid R773 encodes an anion pump. The pump has two polypeptide components. The catalytic subunit, the ArsA protein, is an oxyanion-stimulated ATPase. The membrane component, the ArsB protein, has been localized in the inner membrane of Escherichia coli. The ArsA and ArsB proteins have been postulated to form a membrane complex which functions as an anion-translocating ATPase. In this study evidence is presented showing that expression of the arsB gene is required to anchor the ArsA protein to the inner membrane. Binding studies with purified ArsA to membranes with and without the arsB gene product confirm this requirement. Membranes of uncA mutants containing both the ArsA and ArsB proteins exhibit arsenite(antimonite)-stimulated ATPase activity. These results support the model in which the ArsA protein is the catalytic energy transducing component of the anion pump, whereas the integral membrane ArsB protein serves as both the anion channel and membrane binding site for the ArsA protein.
Tisa
L S
LS
Department of Biochemistry, Wayne State University, School of Medicine, Detroit, Michigan 48201.
Rosen
B P
BP
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D000838:Anions; D000965:Antimony / Q000494:pharmacology; D001151:Arsenic / Q000494:pharmacology; D053501:Arsenite Transporting ATPases; D018053:Arsenites; D001426:Bacterial Proteins / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D004352:Drug Resistance, Microbial / Q000235:genetics*; D004926:Escherichia coli / Q000378:metabolism*; D015870:Gene Expression; D005798:Genes, Bacterial; D015151:Immunoblotting; D007473:Ion Channels / Q000378:metabolism*; D016623:Ion Pumps; D008565:Membrane Proteins / Q000378:metabolism; D009097:Multienzyme Complexes; D009876:Operon; D010957:Plasmids
D000838:Anions; D018053:Arsenites; D001426:Bacterial Proteins; D007473:Ion Channels; D016623:Ion Pumps; D008565:Membrane Proteins; D009097:Multienzyme Complexes; C061374:antimonite; D000965:Antimony; D000251:Adenosine Triphosphatases; D053501:Arsenite Transporting ATPases; C015001:arsenite; D001151:Arsenic