Bacterial lactoferrin receptors: insights from characterizing the Moraxella bovis receptors.
Moraxella bovis is the causative agent of infectious conjunctivitis in cattle. Moraxella bovis isolates were shown to specifically bind bovine lactoferrin (bLf) and bovine transferrin (bTf) and to use these proteins as a source of iron to support the growth of iron-limited cells. Affinity isolation experiments with immobilized bTf yielded two proteins readily resolved by SDS-PAGE analysis, whereas only a single band of approximately 100 kDa was detected when immobilized bLf was used as the affinity ligand. Using a novel cloning strategy, regions containing the genes encoding the lactoferrin (Lf) and transferrin (Tf) receptor proteins were isolated and sequenced, demonstrating that they both consisted of two genes, with the tbpB or lbpB gene preceding the tbpA or lbpA gene. The cloned lbp genes were used to generate isogenic mutants deficient in lactoferrin binding protein A and (or) B, and the resulting strains were tested in growth and binding assays. The isogenic mutants were deficient in their use of bLf for growth and had substantially diminished bLf binding capability. The predicted amino acid sequence from the segment encoding Lf binding protein B revealed an internal amino acid homology suggesting it is a bi-lobed protein, with a C-lobe enriched in acidic amino acids, but without the evident clustering observed in Lf-binding proteins from other species.
Yu
Rong-Hua
RH
Department of Microbiology and Infectious Diseases, University of Calgary, Alberta, Canada.
Schryvers
Anthony B
AB
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D001428:Bacterial Vaccines / Q000276:immunology; D002417:Cattle; D002455:Cell Division; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D005798:Genes, Bacterial / Q000235:genetics; D007501:Iron / Q000378:metabolism; D007781:Lactoferrin / Q000378:metabolism; D008024:Ligands; D008969:Molecular Sequence Data; D016955:Moraxella bovis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D013379:Substrate Specificity; D014168:Transferrin / Q000378:metabolism
D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D008024:Ligands; D011956:Receptors, Cell Surface; D014168:Transferrin; C038340:lactoferrin receptors; D007501:Iron; D007781:Lactoferrin