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.
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June 2002
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Research Article|
June 15 2002
Expression and purification of functional recombinant meningococcal transferrin-binding protein A
Jonathan S. OAKHILL;
Jonathan S. OAKHILL
∗Metalloprotein Research Group, The Randall Centre for Molecular Mechanisms of Cell Function, King's College London, London SE1 1UL, U.K.
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Christopher L. JOANNOU;
Christopher L. JOANNOU
∗Metalloprotein Research Group, The Randall Centre for Molecular Mechanisms of Cell Function, King's College London, London SE1 1UL, U.K.
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Susan K. BUCHANAN;
Susan K. BUCHANAN
1
†School of Crystallography, Birkbeck College, Malet Street, London WC1E 7HX, U.K.
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Andrew R. GORRINGE;
Andrew R. GORRINGE
‡Centre for Applied Microbiology and Research, Porton Down, Salisbury SP4 0JG, Wiltshire, U.K.
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Robert W. EVANS
Robert W. EVANS
2
∗Metalloprotein Research Group, The Randall Centre for Molecular Mechanisms of Cell Function, King's College London, London SE1 1UL, U.K.
2To whom correspondence should be addressed (e-mail robert.evans@kcl.ac.uk).
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Publisher: Portland Press Ltd
Received:
March 28 2002
Revision Received:
April 23 2002
Accepted:
April 25 2002
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2002
2002
Biochem J (2002) 364 (3): 613–616.
Article history
Received:
March 28 2002
Revision Received:
April 23 2002
Accepted:
April 25 2002
Citation
Jonathan S. OAKHILL, Christopher L. JOANNOU, Susan K. BUCHANAN, Andrew R. GORRINGE, Robert W. EVANS; Expression and purification of functional recombinant meningococcal transferrin-binding protein A. Biochem J 15 June 2002; 364 (3): 613–616. doi: https://doi.org/10.1042/bj20020500
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