Molecular and immunological analysis of a fibronectin-binding protein antigen secreted by Mycobacterium Ieprae

J. E R Thole, R. Schöningh, A. A M Janson, T. Garbe, Y. E. Cornelisse, J. E. Clark-Curtiss, A. H J Kolk, T. H M Ottenhoff, R. R P De Vries, C. Abou-Zeid

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Abstract

By screening a Mycobacterium leprae λgt11 genomic DNA library with leprosy-patient sera we have previously identified 50 recombinant clones that expressed novel M. leprae antigens (Sathish et al., 1990). In this study, we show by DNA sequencing and immunoblot analysis that three of these clones express a M. leprae homologue of the fibronectin-binding antigen 85 complex of mycobacteria. The complete gene was characterized and it encodes a 327-amino-acid polypeptide, consisting of a consensus signal sequence of 38 amino acids followed by a mature protein of 289 amino acids. This is the first sequence of a member of the M. leprae antigen 85 complex, and Southern blotting analysis indicated the presence of multiple genes of the 85 complex in the genome of M. leprae. The amino acid sequence displays 75-85% sequence identity with components of the antigen 85 complex from M. tuberculosis, M. bovis BCG and M. kansasii. Furthermore, antibodies to the antigen 85 complex of M. tuberculosis and M. bovis BCG reacted with two fusion proteins containing the amino acid regions 55-266 and 265-327 of the Af. leprae protein. The M. leprae 30/31 kDa protein induces strong humoral and cellular responses, as judged by Western blot analysis with patient sera and proliferation of T cells derived from healthy individuals and leprosy patients. Amino acid regions 55-266 and 265-327 both were shown to bind to fibronectin, indicating the presence of at least two fibronectin-binding sites on the M. leprae protein. These data indicate that this 30/31 kDa protein is not only important in the immune response against M. leprae, but may also have a biological role in the interaction of this bacillus with the human host.

Original languageEnglish (US)
Pages (from-to)153-163
Number of pages11
JournalMolecular Microbiology
Volume6
Issue number2
StatePublished - Jan 1992
Externally publishedYes

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Mycobacterium leprae
Mycobacterium
Fibronectins
Carrier Proteins
Antigens
Amino Acids
Leprosy
Mycobacterium bovis
Proteins
Tuberculosis
Clone Cells
Genomic Library
Consensus Sequence
Protein Sorting Signals
Southern Blotting
Serum
Gene Library
DNA Sequence Analysis
Bacillus
Genes

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Thole, J. E. R., Schöningh, R., Janson, A. A. M., Garbe, T., Cornelisse, Y. E., Clark-Curtiss, J. E., ... Abou-Zeid, C. (1992). Molecular and immunological analysis of a fibronectin-binding protein antigen secreted by Mycobacterium Ieprae. Molecular Microbiology, 6(2), 153-163.

Molecular and immunological analysis of a fibronectin-binding protein antigen secreted by Mycobacterium Ieprae. / Thole, J. E R; Schöningh, R.; Janson, A. A M; Garbe, T.; Cornelisse, Y. E.; Clark-Curtiss, J. E.; Kolk, A. H J; Ottenhoff, T. H M; De Vries, R. R P; Abou-Zeid, C.

In: Molecular Microbiology, Vol. 6, No. 2, 01.1992, p. 153-163.

Research output: Contribution to journalArticle

Thole, JER, Schöningh, R, Janson, AAM, Garbe, T, Cornelisse, YE, Clark-Curtiss, JE, Kolk, AHJ, Ottenhoff, THM, De Vries, RRP & Abou-Zeid, C 1992, 'Molecular and immunological analysis of a fibronectin-binding protein antigen secreted by Mycobacterium Ieprae', Molecular Microbiology, vol. 6, no. 2, pp. 153-163.
Thole JER, Schöningh R, Janson AAM, Garbe T, Cornelisse YE, Clark-Curtiss JE et al. Molecular and immunological analysis of a fibronectin-binding protein antigen secreted by Mycobacterium Ieprae. Molecular Microbiology. 1992 Jan;6(2):153-163.
Thole, J. E R ; Schöningh, R. ; Janson, A. A M ; Garbe, T. ; Cornelisse, Y. E. ; Clark-Curtiss, J. E. ; Kolk, A. H J ; Ottenhoff, T. H M ; De Vries, R. R P ; Abou-Zeid, C. / Molecular and immunological analysis of a fibronectin-binding protein antigen secreted by Mycobacterium Ieprae. In: Molecular Microbiology. 1992 ; Vol. 6, No. 2. pp. 153-163.
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