title: Expression and purification of functional recombinant meningococcal transferrin-binding protein A.
abstract: Pathogenic bacteria of the genus Neisseria have a siderophore-independent iron-uptake system reliant on a direct interaction between the bacterial cell and human transferrin (hTf), a serum protein. In the meningococcus, this uptake system is dependent on two surface-exposed, transferrin-binding proteins (Tbps), TbpA and TbpB. TbpA is highly conserved among meningococcal strains, and is thought to be a porin-like integral protein that functions as a gated channel for the passage of iron into the periplasm. TbpB is more variable in size, lipidated and fully surface-exposed. Given its location on the cell surface, its role in pathogenicity and interstrain sequence conservation, TbpA is currently being regarded for inclusion in a meningococcal vaccine effective against all serogroups. This requires gaining knowledge of the ligand-receptor interactions. In the present study we have optimized a procedure for obtaining purified, functionally active recombinant TbpA at a level and stability necessary for the initiation of such studies.
authors.affiliation: Metalloprotein Research Group, The Randall Centre for Molecular Mechanisms of Cell Function, King's College London, London SE1 1UL, UK.
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Oakhill
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Joannou
authors.forename: Susan K
authors.initials: SK
authors.lastname: Buchanan
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Gorringe
authors.forename: Robert W
authors.initials: RW
authors.lastname: Evans
mesh_terms: D002352:Carrier Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D033862:Iron-Binding Proteins; D009345:Neisseria meningitidis / Q000235:genetics*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D014168:Transferrin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D033863:Transferrin-Binding Proteins
chemical_list: D002352:Carrier Proteins; D033862:Iron-Binding Proteins; D011994:Recombinant Proteins; D014168:Transferrin; D033863:Transferrin-Binding Proteins