The bacterial receptor protein, transferrin-binding protein B, does not independently facilitate the release of metal ion from human transferrin.
Pathogenic Gram-negative bacteria of the Pasteurellaceae and Neisseriaceae acquire iron for growth from host transferrin through the action of specific surface receptors. Iron is removed from transferrin by the receptor at the cell surface and is transported across the outer membrane to the periplasm. A periplasmic binding protein-dependent pathway subsequently transports iron into the cell. The transferrin receptor is composed of a largely surface-exposed lipoprotein, transferrin binding protein B, and a TonB-dependent integral outer membrane protein, transferrin binding protein A. To examine the role of transferrin binding protein B in the iron removal process, complexes of recombinant transferrin binding protein B and transferrin were prepared and compared with transferrin in metal-binding and -removal experiments. A polyhistidine-tagged form of recombinant transferrin binding protein B was able to purify a complex with transferrin that was largely monodisperse by dynamic light scattering analysis. Gallium was used instead of iron in the metal-binding studies, since it resulted in increased stability of recombinant transferrin binding protein B in the complex. Difference absorption spectra were used to monitor removal of gallium by nitrilotriacetic acid. Kinetic and equilibrium binding studies indicated that transferrin binds gallium more tightly in the presence of transferrin binding protein B. Thus, transferrin binding protein B does not facilitate metal ion removal and additional components are required for this process.
Department of Biological Sciences, Faculty of Medicine, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada.
Ulyana
U
Nemish
Rong-Hua
RH
Yu
Leslie W
LW
Tari
Karla
K
Krewulak
Anthony B
AB
Schryvers
D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001692:Biological Transport; D005708:Gallium / Q000378:metabolism; D006801:Humans; D007501:Iron / Q000378:metabolism*; D009345:Neisseria meningitidis / Q000378:metabolism; D019897:Periplasm / Q000378:metabolism; D011485:Protein Binding; D011990:Receptors, Transferrin / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism*; D033882:Transferrin-Binding Protein B / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D011990:Receptors, Transferrin; D011994:Recombinant Proteins; D014168:Transferrin; D033882:Transferrin-Binding Protein B; D005708:Gallium; D007501:Iron