Interaction of the catalytic and the membrane subunits of an oxyanion-translocating ATPase.
Resistance to arsenical and antimonial compounds in Escherichia coli is due to active extrusion of these compounds from cells expressing the ars operon. The arsenical pump is an ion-translocating ATPase which consists of two polypeptide components, the ArsA and ArsB proteins. The ArsB protein, the inner membrane component of the pump, has been shown to function as the membrane anchor for the catalytic subunit, the ArsA protein. The properties and nature of interaction between these two components of the pump were investigated using an in vitro binding assay. Purified ArsA protein bound to the membrane in a saturable manner. In the absence of arsenite or antimonite an apparent positive cooperativity in the binding of the ArsA protein to membrane vesicles containing the ArsB protein was observed. In the presence of arsenite or antimonite binding became hyperbolic, with a 10-fold decrease in the concentration of ArsA protein required for half-maximal binding, without any change in the stoichiometry of the complex. Addition of ATP had little affect on membrane binding of the ArsA ATPase subunit. In the presence or absence of the anionic substrates binding was maximal in a pH range 7.5-8.5.
Dey
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Department of Biochemistry, Wayne State University, School of Medicine, Detroit, Michigan 48201.
Dou
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Tisa
L S
LS
Rosen
B P
BP
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000965:Antimony / Q000633:toxicity; D020261:Arsenic Poisoning; D053501:Arsenite Transporting ATPases; D002462:Cell Membrane / Q000201:enzymology; D004352:Drug Resistance, Microbial; D004790:Enzyme Induction; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D005838:Genotype; D006863:Hydrogen-Ion Concentration; D016623:Ion Pumps; D007700:Kinetics; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000378:metabolism; D009097:Multienzyme Complexes; D009876:Operon; D011485:Protein Binding
D016623:Ion Pumps; D046911:Macromolecular Substances; D008565:Membrane Proteins; D009097:Multienzyme Complexes; D000965:Antimony; D000251:Adenosine Triphosphatases; D053501:Arsenite Transporting ATPases