Transferrin binding protein two interacts with both the N-lobe and C-lobe of ovotransferrin.
The present study was initiated to identify the region(s) of ovotransferrin involved in binding to the bacterial transferrin receptors from Haemophilus paragallinarum and Haemophilus avium. Ovotransferrin was digested with either trypsin or thermolysin to obtain its N-lobe and C-lobe fragments. The individual fragments were then purified by a combination of gel exclusion and ion-exchange chromatography. Solid phase binding experiments with the individual fragments demonstrated that the C-lobe fragments blocked the binding of horse radish peroxidase-conjugated ovotransferrin to the transferrin receptors and that much higher concentrations of the N-lobe fragment were required for any detectable blocking. Affinity isolation of the bacterial transferrin receptor from the two Haemophilus species revealed that both native ovotransferrin and its C-lobe fragment were capable of isolating two iron repressible outer membrane proteins. These 95 and 60 kDa outer membrane proteins correspond to Tbp1 and Tpb2, respectively. In contrast, the N-lobe fragment was capable of isolating Tbp2 of H. paragallinarum but not that of H. avium. The inability of the N-lobe and C-lobe fragments from ovotransferrin and human transferrin to support the growth of iron-limited cultures of H. paragallinarum and Neisseria meningitidis, respectively, suggested that interaction with both lobes is necessary for efficient iron acquisition.
Alcantara
J
J
Department of Microbiology and Infectious Diseases, University of Calgary, Alberta, Canada.
Schryvers
A B
AB
D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism* / Q000502:physiology; D002352:Carrier Proteins / Q000378:metabolism / Q000502:physiology; D003207:Conalbumin / Q000737:chemistry* / Q000378:metabolism; D006190:Haemophilus / Q000737:chemistry / Q000254:growth & development / Q000378:metabolism; D007501:Iron / Q000378:metabolism*; D033862:Iron-Binding Proteins; D009345:Neisseria meningitidis / Q000254:growth & development / Q000378:metabolism; D010446:Peptide Fragments / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology; D011485:Protein Binding; D011990:Receptors, Transferrin / Q000302:isolation & purification / Q000378:metabolism* / Q000502:physiology; D033863:Transferrin-Binding Proteins
D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D033862:Iron-Binding Proteins; D010446:Peptide Fragments; D011990:Receptors, Transferrin; D033863:Transferrin-Binding Proteins; D003207:Conalbumin; D007501:Iron