Pyruvylated glycolipids from Mycobacterium smegmatis. Nature and location of the lipid components

The dipyruvylated glycolipid from Mycobacterium smegmatis (Saadat, S., and Ballou, C.E. (1983) J. Biol. Chem. 258, 1813-1818) has been shown to have the following structure in which FA1 is tetra- or hexadecanoic acid and FA2 is 2,4-dimethyl-2-eicosenoic acid. (formula; see text) The fast atom bombardment mass spectrum showed two major ions [M - H]- at m/z 1511 and 1539 (Mr 1512 and 1540) in a ratio of 1.4:1, suggesting that the glycolipid was a mixture of homologs that differed in fatty acid composition by 2 methylene groups. Analysis revealed C14, C16, and C22 fatty acids in ratios of 0.6:0.4:1.0, indicating that 60% of the molecules contained a C14 and C22 fatty acid whereas 40% contained a C16 and C22 fatty acid. The fragmentation pattern showed that a single glucose unit along with the smaller fatty acid could be lost to yield a tetrasaccharide with attached C22 fatty acid, and a second fragmentation yielded a trisaccharide containing 2 pyruvic acids but without attached fatty acid. The C14 and C16 fatty acids were identified as myristic and palmitic acid, whereas the C22 fatty acid was 2,4-dimethyl-2-eicosenoic acid. Precise localization of the fatty acids came from periodate oxidation and methylation analysis.