title: Response of Haemophilus somnus to iron limitation: expression and identification of a bovine-specific transferrin receptor.
abstract: Nine clinical isolates of Haemophilus somnus were screened for their ability to use different transferrins as a source of iron growth. All nine strains were capable of using bovine but not porcine, human or chicken transferrin. A screening assay for siderophore production did not show any evidence of siderophore production by these strains. When iron-deficient cells from these strains were screened for their ability to bind peroxidase-conjugated transferrin, binding was detected with conjugated bovine, but not human or porcine transferrin. Competition binding studies demonstrated that the binding of peroxidase-conjugated bovine transferrin was competitively inhibited by unconjugated bovine transferrin but not transferrin from other species. The induction of receptor expression by low iron conditions was inhibited by chloramphenicol and rifampicin but not ampicillin indicating that new protein and mRNA synthesis was required for expression of receptor activity. Affinity isolation of receptor proteins with biotinylated bovine transferrin, but not human or porcine transferrin, yielded three proteins from H. somnus strain H74. Two of the proteins were identified as 105 kDa and 73 kDa iron-regulated outer membrane proteins. A third protein of 85 kDa that was isolated did not co-migrate with any iron-regulated outer membrane protein. Affinity isolation of receptor proteins from other strains of H. somnus yielded a 73 kDa protein from all strains and a 105 kDa and 85 kDa protein in four of the six strains analysed.
authors: Department of Microbiology and Infectious Diseases, University of Calgary, Alberta, Canada.
authors: J A
authors: JA
authors: Ogunnariwo
authors: C
authors: C
authors: Cheng
authors: J
authors: J
authors: Ford
authors: A B
authors: AB
authors: Schryvers
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D002417:Cattle; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D006190:Haemophilus / Q000254:growth & development / Q000378:metabolism*; D006801:Humans; D007501:Iron / Q000378:metabolism*; D007502:Iron Chelating Agents / Q000378:metabolism; D011990:Receptors, Transferrin / Q000302:isolation & purification / Q000378:metabolism*; D017262:Siderophores; D013045:Species Specificity; D014168:Transferrin / Q000378:metabolism*
chemical_list: D007502:Iron Chelating Agents; D011990:Receptors, Transferrin; D017262:Siderophores; D014168:Transferrin; D007501:Iron