1887

Abstract

The activity of the two almost identical K-uptake systems, Trk and Trk, from K-12 depends completely and partially on the presence of the gene, respectively. maps inside the operon, which encodes an ATP-binding cassette (ABC) transporter of unknown function from the subgroup of peptide-uptake systems. This study was carried out to clarify the role of gene products in the ATP dependence of the Trk systems. For this purpose Δ Δ and Δ Δ strains of containing plasmids with genes from either or were used. All five plasmid-encoded Sap proteins were made in mini-cells. The presence of the ATP-binding SapD protein from either or alone was sufficient for stimulating the K transport activity of the Trk and Trk systems. K-uptake experiments with cells containing SapD variants with changes in the Walker A box Lys-46 residue, the Walker B box Asp-183 residue and the signature motif residues Gly-162 or Gln-165 suggested that adenine nucleotide binding to SapD rather than ATP hydrolysis by this subunit is required for the activity of the Trk system. K transport via two plasmid-encoded Trk systems in a Δ strain remained dependent on both a high membrane potential and a high cytoplasmic ATP concentration, indicating that in ATP dependence of Trk activity can be independent of Sap proteins. These data are interpreted to mean that Trk systems can interact with an ABC protein other than SapD.

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2001-11-01
2024-04-16
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