In vitro assembly of an anion-stimulated ATPase from peptide fragments.
The oxyanion-translocating ATPase encoded by the ars operon of plasmid R 773 confers resistance to antimonials and arsenicals in Escherichia coli by extrusion of the oxyanions from the cells. The catalytic subunit, the ArsA protein, is an oxyanion-stimulated ATPase with two nucleotide binding consensus sequences, one in the N-terminal half and one in the C-terminal half of the protein. In this report subclones of the arsA gene were constructed to produce polypeptide fragments of the ArsA protein. By themselves none of the fragments exhibited anion-stimulated ATPase activity. Denaturation and renaturation of mixtures of N- and C-terminal polypeptides that between them comprised an entire ArsA protein resulted in active ATPase complexes.
Kaur
P
P
Department of Biochemistry, Wayne State University, School of Medicine, Detroit, Michigan 48201.
Rosen
B P
BP
D000251:Adenosine Triphosphatases / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D053501:Arsenite Transporting ATPases; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D016623:Ion Pumps; D008565:Membrane Proteins / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D009876:Operon; D010446:Peptide Fragments / Q000378:metabolism; D011489:Protein Denaturation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism
D016623:Ion Pumps; D008565:Membrane Proteins; D009097:Multienzyme Complexes; D010446:Peptide Fragments; D011994:Recombinant Proteins; D000251:Adenosine Triphosphatases; D053501:Arsenite Transporting ATPases