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Immunodominant B‐Cell Epitope in the Mce1A Mammalian Cell Entry Protein of Mycobacterium tuberculosis Cross‐Reacting with Glutathione S‐Transferase
Author(s) -
Harboe M.,
Das A. K.,
Mitra D.,
Ulvund G.,
Ahmad S.,
Harkness R. E.,
Das D.,
Mustafa A. S.,
Wiker H. G.
Publication year - 2004
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.0300-9475.2004.01383.x
Subject(s) - epitope , monoclonal antibody , polyclonal antibodies , microbiology and biotechnology , mycobacterium tuberculosis , antibody , chemistry , epitope mapping , linear epitope , monoclonal , biology , tuberculosis , immunology , medicine , pathology
The TB1‐5 76C monoclonal antibody raised against a synthetic 60‐mer peptide in the N‐terminal part of the Mce1A mammalian cell entry protein of Mycobacterium tuberculosis has previously been shown to react with a linear epitope in the KRRITPKD region, residues 131–138 in Mce1A, and to cross‐react with Mce1F. Six additional monoclonal antibodies raised against the same peptide were also shown to cross‐react with Mce1F. Four of them reacted with a linear epitope in the same area, indicating that this area is immunodominant but showed distinct differrences in fine specificity. Two monoclonal antibodies did not react with synthetic peptides from this region on the solid phase in enzyme‐linked immunosorbent assay, indicating greater influence of conformation on reactivity. None of the monoclonal antibodies reacted with 14‐mer synthetic peptides from the corresponding area in Mce2A, Mce3A, Mce4A, M. avium , M. smegmatis or M. leprae . The reaction pattern of the monoclonal antibodies was analysed in relation to our model of the Mce1A molecule (AK Das et al . Biochem Biophys Res Commun 2003;302:442–7).The epitope is located on the surface of Mce1A, at the distal β‐strand‐loop region in the β‐domain supporting its potential role in promoting uptake of M. tuberculosis in host cells. Monoclonal antibody TB1‐5 19C cross‐reacted with glutathione S‐transferase of Schistosoma japonicum containing a PKE triplet. Monoclonal antibody TB1‐5 76C gave a major band at about 44 kDa in Western blotting of M. tuberculosis sonicate, whereas polyclonal rabbit anti‐Mce1A peptide antibodies reacting with the extended TTPKNPTKRRITPKDVI area of Mce1A showed a distinct band above the 160 kDa molecular mass standard.