The domains of human fibronectin mediating the binding of alpha antigen, the most immunopotent antigen of mycobacteria that induces protective immunity against mycobacterial infection.
We have recently shown that alpha antigen (alpha-Ag), the immunodominant antigen of mycobacteria, has a novel fibronectin (FN)-binding motif that is unique among mycobacteria [Naito, Ohara, Matsumoto and Yamada (1998) J. Biol. Chem. 273, 2905-2909]. In this study, we examined the domains of human FN that interacted with alpha-Ag. Fragments of FN generated by either proteolysis or recombinant DNA techniques were compared for their ability to bind to alpha-Ag. Fragments containing either the C-terminal heparin-binding domain or the central cell-binding domain consistently bound to alpha-Ag. The fragment of the C-terminal heparin-binding domain, upon mutation that resulted in the loss of its heparin-binding activity, could not bind with alpha-Ag. These findings suggested that the mutated site, i.e. the main heparin-binding site of FN, was also the principal site for binding to alpha-Ag. The alpha-Ag-binding domains of FN could bind whole mycobacterial bacilli, suggesting that these two domains are important contributors to mycobacterial infection.
Naito
M
M
Department of Oral Bacteriology, Nagasaki University School of Dentistry, Sakamoto 1-7-1, Nagasaki 852-8588, Japan. mnaito@net.nagasaki-u.ac.jp
Fukuda
T
T
Sekiguchi
K
K
Yamada
T
T
D000942:Antigens, Bacterial / Q000276:immunology* / Q000378:metabolism*; D001667:Binding, Competitive; D019921:Biotinylation; D004797:Enzyme-Linked Immunosorbent Assay; D005353:Fibronectins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005780:Gelatin / Q000378:metabolism; D006493:Heparin / Q000378:metabolism; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology / Q000378:metabolism; D008970:Molecular Weight; D009154:Mutation / Q000235:genetics; D009164:Mycobacterium Infections / Q000276:immunology*; D009163:Mycobacterium bovis / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013820:Thermolysin / Q000378:metabolism
D000942:Antigens, Bacterial; D005353:Fibronectins; D016056:Immunodominant Epitopes; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C052242:alpha antigens, Mycobacterium; D005780:Gelatin; D006493:Heparin; D013820:Thermolysin