Skin reactivity and fibronectin-binding property of TB66 (66-kDa protein of Mycobacterium tuberculosis).
A 66-kDa protein (TB66) was purified from culture filtrate (CF) and cell sonicate (CS) of Mycobacterium tuberculosis H37Rv by immobilised metal affinity chromatography (IMAC) on a Ni-nitrilotriacetic acid (NTA) column. TB66 was found to be a fibronectin-binding protein as determined by ELISA and could be purified by affinity chromatography with fibronectin-Sepharose. A similar 66-kDa protein could be isolated also from M. bovis, M. bovis BCG, M. africanum and M. tuberculosis H37Ra by IMAC, but not from any other mycobacteria. The NH2-terminal amino-acid sequence of TB66 from H37Rv and M. bovis was identical and showed 85% homology with the N-terminal sequence of bovine serum albumin (BSA). A monoclonal antibody (MAb) OD4AG3 recognised a heat-stable and trypsin-sensitive epitope near the C-terminal end of TB66. This MAb also recognised the 66-kDa protein isolated from the other members of the M. tuberculosis complex. In tests of immunogenicity, TB66 elicited a delayed type hypersensitivity reaction in guinea-pigs immunised with either TB66 or with M. tuberculosis H37Rv. TB66 also elicited an antibody response in immunised guinea-pigs and stimulated murine macrophages to produce tumour necrosis factor.
Department of Microbiology and Immunology, Morehouse School of Medicine, Atlanta, Georgia 30310.
R G
RG
Deshpande
M B
MB
Khan
D A
DA
Bhat
R G
RG
Navalkar
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D005353:Fibronectins / Q000378:metabolism*; D006168:Guinea Pigs; D006968:Hypersensitivity, Delayed; D015151:Immunoblotting; D008264:Macrophages / Q000276:immunology; D008297:Male; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000737:chemistry* / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D002352:Carrier Proteins; D005353:Fibronectins; C090440:TB66 protein, Mycobacterium tuberculosis; D014409:Tumor Necrosis Factor-alpha