PMID:8762140
| Citation |
Barroga, CF, Zhang, H, Wajih, N, Bouyer, JH and Hermodson, MA (1996) The proteins encoded by the rbs operon of Escherichia coli: I. Overproduction, purification, characterization, and functional analysis of RbsA. Protein Sci. 5:1093-9 |
|---|---|
| Abstract |
The nucleotide-binding component of the high-affinity ribose transport system of Escherichia coli, RbsA, was overproduced from a T7-7 expression vector, and the protein was purified. Biochemical analyses of the purified protein indicated that the ATP analogues, 5'-FSBA and 8-azido ATP, covalently labeled the protein, a reaction that was inhibited by ATP, but not by GTP or CTP. The pure protein exhibited low-level ATPase activity with a K(m) of about 140 microM. Analyses of bacterial strains carrying chromosomal deletions of rbsA and other rbs genes suggested that RbsA is important for the chemotaxis function, a surprising result that was not anticipated from previous studies. However, an inconsistency between the several results from deletion strains raises questions regarding the interpretations of the in vivo data. |
| Links |
PubMed PMC2143435 Online version:10.1002/pro.5560050611 |
| Keywords |
ATP-Binding Cassette Transporters/genetics; ATP-Binding Cassette Transporters/isolation & purification; ATP-Binding Cassette Transporters/metabolism; Adenosine/analogs & derivatives; Adenosine/chemistry; Adenosine Triphosphatases/metabolism; Adenosine Triphosphate/analogs & derivatives; Adenosine Triphosphate/analysis; Adenosine Triphosphate/chemistry; Adenosine Triphosphate/metabolism; Affinity Labels/analysis; Affinity Labels/chemistry; Animals; Autoradiography; Azides/analysis; Azides/chemistry; Blotting, Western; Carrier Proteins/genetics; Carrier Proteins/immunology; Carrier Proteins/isolation & purification; Carrier Proteins/metabolism; Chemotaxis; Chromosome Deletion; DNA, Bacterial/genetics; Electrophoresis, Polyacrylamide Gel; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli Proteins; Gene Deletion; Gene Expression/genetics; Genetic Vectors; Membrane Transport Proteins; Operon; Photolysis; Plasmids; Rabbits; Recombinant Proteins/genetics; Recombinant Proteins/isolation & purification; Recombinant Proteins/metabolism |
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