SUMMARY: Internal radiolabelling procedures were used to radiolabel the oligosaccharide determinant of the glycopeptidolipids (GPL) from serovars 4 and 20 of the complex. Mycobacteria were cultured in the presence of [6-H]fucose, [2-H]mannose or [-H]methionine, after which radiolabeled native lipid was extracted and distribution of radioactivity in native and deacetylated lipid was determined by thin-layer chromatographic methods. Incorporation of radiolabel was confirmed by examining acid hydrolysates of purified GPL for H-labelled sugars on cellulose thin-layer plates. Least incorporation of radiolabel into GPL was observed with [6-H]fucose, whereas better incorporation was obtained with [2-H]mannose and [-H]methiomne. Use of [-H]methiomne resulted in the radiolabelling of the methylated sugars in both the oligosaccharide determinant and the 3,4-di--methylrhamnose located at the terminus of the peptide core. Use of [2-H]mannose resulted in the incorporation of radioactivity into the oligosaccharide determinant as 2--methylfucose, found in the GPL of both serovars 4 and 20. GPL radiolabeled with [2-H]mannose were subsequently examined in macrophage cultures and found to be relatively inert to degradation by those phagocytic cells. These results substantiate earlier findings with the GPL of serovar 20 and indicate that these mycobacterial components may play a role in pathogenesis.


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