Purification and characterization of a tuberculin-active substance from Mycobacterium bovis BCG.
A new tuberculin-active substance, designated TAS-1D3, has been purified from the extract of Mycobacterium bovis BCG by precipitation at pH 4.2, ethanol fractionation, and column chromatography involving CM-cellulose, QAE-Sephadex A-25, Sephadex G-100, and Sephadex G-75. TAS-1D3 was homogeneous in polyacrylamide gel electrophoresis and positive in both Coomassie brilliant blue and periodic acid-Shiff staining, suggesting that TAS-1D3 is a glycoprotein. The molecular weight of TAS-1D3 was estimated to be 26,000 by gel filtration. In amino acid analysis, TAS-1D3 was distinctive in having proline as a dominant amino acid, and in that it lacked basic amino acids, sulfur-containing amino acids and aromatic amino acids. Moreover, TAS-1D3 was almost devoid of absorption at around 280 nm. In guinea pigs sensitized with BCG vaccine, the tuberculin activity of TAS-1D3 was about forty times more potent than that of purified protein derivative (PPD).
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001426:Bacterial Proteins; D002241:Carbohydrates / Q000032:analysis; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000302:isolation & purification; D008970:Molecular Weight; D009163:Mycobacterium bovis / Q000032:analysis*; D010455:Peptides / Q000008:administration & dosage / Q000302:isolation & purification*; D012882:Skin Tests; D014373:Tuberculin / Q000008:administration & dosage / Q000302:isolation & purification*
D000596:Amino Acids; D001426:Bacterial Proteins; D002241:Carbohydrates; D006023:Glycoproteins; D010455:Peptides; D014373:Tuberculin; C009610:tuberculin-active protein, Mycobacterium