Renshaw, P.S. and Panagiotidou, P. and Whelan, A. and Gordon, S.V. and Hewinson, R.G. and Williamson, R.A. and Carr, M.D. (2002) Conclusive evidence that the major T-cell antigens of the Mycobacterium tuberculosis complex ESAT-6 and CFP-10 form a tight, 1:1 complex and characterization of the structural properties of ESAT-6, CFP-10, and the ESAT-6*CFP-10 complex. Implications for pathogenesis and virulence. Journal of Biological Chemistry, 277 (24). pp. 21598-21603. ISSN 0021-9258 .
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The proteins ESAT-6 and CFP-10 have been shown to be secreted by Mycobacterium tuberculosis and Mycobacterium bovis cells, to be potent T-cell antigens, and to have a clear but as yet undefined role in tuberculosis pathogenesis. We have successfully overexpressed both ESAT-6 and CFP-10 in Escherichia coli and developed efficient purification schemes. Under in vivo-like conditions, a combination of fluorescence, circular dichroism, and nuclear magnetic resonance spectroscopy have shown that ESAT-6 contains up to 75% helical secondary structure, but little if any stable tertiary structure, and exists in a molten globule-like state. In contrast, CFP-10 was found to form an unstructured, random coil polypeptide. An exciting discovery was that ESAT-6 and CFP-10 form a tight, 1:1 complex, in which both proteins adopt a fully folded structure, with about two-thirds of the backbone in a regular helical conformation. This clearly suggests that ESAT-6 and CFP-10 are active as the complex and raises the interesting question of whether other ESAT-6/CFP-10 family proteins (22 paired genes in M. tuberculosis) also form tight, 1:1 complexes, and if so, is this limited to their genome partner, or is there scope for wider interactions within the protein family, which could provide greater functional flexibility?
|Additional information:||0021-9258 (Print) Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.|
|Uncontrolled keywords:||Antigens/metabolism Antigens, Bacterial/chemistry/metabolism Bacterial Proteins/chemistry/metabolism Circular Dichroism Electrophoresis, Polyacrylamide Gel Escherichia coli/metabolism Genetic Vectors Guanidine/pharmacology Histidine/chemistry Magnetic Resonance Spectroscopy Mycobacterium bovis/metabolism Mycobacterium tuberculosis/metabolism Parasympathomimetics/pharmacology Phylogeny Protein Binding Protein Denaturation Protein Structure, Tertiary Spectrometry, Fluorescence T-Lymphocytes/*immunology/metabolism Ultraviolet Rays|
|Divisions:||Faculties > Science Technology and Medical Studies > School of Biosciences > Protein Science Group|
|Depositing User:||Richard Williamson|
|Date Deposited:||09 Sep 2008 16:52|
|Last Modified:||14 Jan 2010 14:20|
|Resource URI:||http://kar.kent.ac.uk/id/eprint/5432 (The current URI for this page, for reference purposes)|
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