Identification, isolation and partial characterization of Mycobacterium tuberculosis glycoprotein antigens.
In Mycobacterium tuberculosis culture filtrates, three concanavalin A (ConA)-binding bands of 55, 50 and 38 kilodaltons (kD) were identified by labelling blotted proteins with a ConA-peroxidase conjugate. Binding was inhibited by the competitor sugar alpha-methyl mannoside and by reduction with sodium m-periodate. Bands of 55, 50 and 38 kD stained with Coomasie blue were sensitive to digestion with proteases, thus indicating that they are proteins. Glycoproteins were isolated by lectin affinity chromatography or by elution from nitrocellulose membranes. On the isolated form, the 55-50 kD doublet glycoprotein was 65.4% protein and 34.6% sugar. The purified 38 kD molecule was 74.3% protein and 25.7% carbohydrate. By immunoblot, antibodies against mycobacterial glycoproteins were demonstrated in immunized rabbits and in patients with pulmonary tuberculosis, but not in healthy individuals. Treatment with sodium m-periodate abolished binding of rabbit antibodies to the 38 kD glycoprotein. Reactivity of the 55-50 kD doublet glycoprotein was not altered by reduction. By immunoblot with monoclonal antibodies TB71 and TB72, a carbohydrate-dependent and a carbohydrate-independent epitope could be identified on the 38 kD glycoprotein.
Espitia
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Departamento de Inmunología, Universidad Nacional Autónoma de México, D.F.
Mancilla
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D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis; D000942:Antigens, Bacterial / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D003208:Concanavalin A / Q000276:immunology; D000939:Epitopes / Q000302:isolation & purification; D006023:Glycoproteins / Q000302:isolation & purification*; D006801:Humans; D008970:Molecular Weight; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D011817:Rabbits; D014397:Tuberculosis, Pulmonary / Q000276:immunology
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D000939:Epitopes; D006023:Glycoproteins; D003208:Concanavalin A