A novel purification of ferric citrate receptor (FecA) from Escherichia coli UT5600 and further characterization of its binding activity.
In our earlier paper, it was demonstrated that the FecA receptor protein from Escherichia coli UT5600/pBB2 (leu-, proC-, trpE-, entA-, rpsl-, delta (ompT-fepA)-/Ampr, fepA) binds with ferric enterobactin. In order to explore this further the outer membrane receptor protein, FecA, has been isolated from UT5600 (fepA-) and purified to homogeneity by DE-52-cellulose anion exchange chromatography followed by MonoPFPLC chromatofocusing. Partially purified FecA and homogeneous FecA show binding activity to [55Fe]ferric enterobactin and the binding is specific. Binding activity of FecA can be enhanced by ferric citrate. Lipopolysaccharide-free FecA as ascertained by silver staining and the endotoxin test still retains the same activity. In vivo uptake studies using different strains of E. coli suggest that FecA in E. coli plays an important role in ferrienterobactin transport.
Zhou
X H
XH
Department of Chemistry and Biochemistry, University of Oklahoma, Norman 73019.
van der Helm
D
D
D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002352:Carrier Proteins; D002852:Chromatography, Ion Exchange; D004758:Enterobactin / Q000378:metabolism; D029968:Escherichia coli Proteins; D005290:Ferric Compounds / Q000378:metabolism; D008070:Lipopolysaccharides / Q000032:analysis; D011485:Protein Binding; D011956:Receptors, Cell Surface; D011955:Receptors, Drug / Q000302:isolation & purification* / Q000378:metabolism; D000091243:Ribosomal Protein S9
D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; C028589:FecA protein, E coli; D005290:Ferric Compounds; D008070:Lipopolysaccharides; D011956:Receptors, Cell Surface; D011955:Receptors, Drug; D000091243:Ribosomal Protein S9; C000717190:RpsI protein, E coli; D004758:Enterobactin; C025314:ferric citrate