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

Renshaw, Philip S. and Panagiotidou, Parthena and Whelan, Adam and Gordon, Stephen V. and Hewinson, R. Glyn and Williamson, Richard A. and Carr, Mark 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 . (The full text of this publication is not available from this repository)

The full text of this publication is not available from this repository. (Contact us about this Publication)
Official URL
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=...

Abstract

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?

Item Type: Article
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
Subjects: Q Science
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: 18 Jun 2014 14:20
Resource URI: http://kar.kent.ac.uk/id/eprint/5432 (The current URI for this page, for reference purposes)
  • Depositors only (login required):