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Bloemink, Marieke J.; Kemmink, Johan; Dentten, Ellen; Muskett, Frederick W.; Whelan, Adam O.; Sheikh, Amina; Hewinson, R. Glyn; Williamson, Richard A.; Carr, Mark D.

doi:10.1023/a:1011239727839

Cited by 2 publications

(2 citation statements)

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“…ments reported for MPB70 previously (9) were readily extended to the 13 C and 1 H side-chain signals using correlations observed in DQF-COSY, TOCSY, TOCSY-HSQC, and HCCH-TOCSY spectra. Apart from exchangeable side-chain groups, the only 1 H signals that remained unidentified were from the backbone amides of G1 and D2 and from the H␥ of the side chain of Leu-119.…”

Section: Resultsmentioning

confidence: 99%

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Solution Structure of the Mycobacterium tuberculosis Complex Protein MPB70

Carr

Bloemink

Dentten

et al. 2003

Journal of Biological Chemistry

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The closely related mycobacteria responsible for tuberculosis produce an unusually high number of secreted proteins, many of which are clearly implicated in pathogenesis and protective immunity. Falling within this category are the closely related proteins MPB70 and MPB83. The structure of MPB70 reveals a complex and novel bacterial fold, which has clear structural homology to the two C-terminal FAS1 domains of the cell adhesion protein fasciclin I, whose structures were reported very recently. Assessment of the surface features of MPB70, the sequence divergence between MPB70 and MPB83, the conservation of residues across a group of FAS1 domains, and the locations of disease-inducing mutations in ␤ig-h3 strongly suggests that MPB70 and MPB83 contain two functional surfaces on opposite faces, which are probably involved in binding to host cell proteins. This analysis also suggests that these functional surfaces are retained in the FAS1 proteins associated with mediating interactions between cells and the extracellular matrix (fasciclin I, periostin, and ␤ig-h3) and furthermore that some of the human corneal disease-inducing substitutions identified in ␤ig-h3 will perturb interactions at these sites.Tuberculosis remains one of the most significant infectious diseases of humans, with about one-third of the world's population currently infected resulting in about 3 million deaths annually (1, 2). The bacteria responsible for tuberculosis belong to the Mycobacterium tuberculosis complex, which is a group of highly related mycobacteria. The complex includes M. tuberculosis, which causes the majority of human tuberculosis, and Mycobacterium bovis, which leads to tuberculosis in a range of domesticated and wild animals. Analysis of the complete M. tuberculosis H37Rv genome has identified the genes for 4006 proteins (3, 4), however, it has so far proved possible to assign specific functions to only about half of the predicted proteins. In addition, we still have relatively little information about which M. tuberculosis complex proteins are essential for pathogenesis, or associated with the stimulation of protective immunity, and even less knowledge of their structures, functions, and mechanisms of action. The finding that only live mycobacterial vaccines provide significant protection against tuberculosis infection (5) clearly indicates a key role for secreted M. tuberculosis complex proteins in stimulating protective immunity and highlights the importance of investigating major secreted antigenic proteins such as MPB70, MPB83, ESAT-6, and CFP-10 (6 -10).The mature M. bovis protein MPB70 and its identical M. tuberculosis homologue MPT70 (Rv2875) are stable, 163-residue polypeptides, that are efficiently secreted from mycobacterial cells following cleavage of a 30-residue signal peptide (3, 6). The proteins contain a single disulfide bond linking Cys-8 and Cys-142 but show no other form of post-translational modification. MPB70 is a major serodominant antigen of M. bovis, which also stimulates cellular immune responses du...

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Section: Resultsmentioning

confidence: 99%

“…15 N-and 15 N/ 13 C-labeled MPB70 were prepared from a pBluescript KS ϩ -based Escherichia coli expression vector (pVW500) as described previously (6,9). The expression system produces the mature form of MPB70 (163 residues) as a secreted product, which is purified from the culture supernatant and periplasmic cell fraction.…”

Section: Sample Preparation-thementioning

confidence: 99%