Affinity purification of mycobacterial polymethyl polysaccharides and a study of polysaccharide-lipid interactions by...

Mycobacterial polymethyl polysaccharides, which bind long-chain fatty acids tightly [Ballou, C.E. (1981) Pure Appl. Chem. 53, 107-112], have been purified on a preparative scale by use of an affinity column packing consisting of (palmitoylamino)alkylsilyl silicate. The relatively large amount of material obtained in this way has allowed a study of the polysaccharide-lipid interactions at millimolar concentrations. The anomeric protons for all of the alpha 1----4-linked hexose units in the mycobacterial methylglucose polysaccharide occur in an envelope centered at delta 5.40, and, on titration with hexadecyltrimethylammonium bromide, the majority of these resonances move upfield to about delta 5.15. This shift is consistent with a change in the polysaccharide from a less ordered chain to one that has a significant proportion of helical conformation, and it is probable that the alkyl chain is included in the coiled portion of the polysaccharide in a manner analogous to the interaction of methylmannose polysaccharide with palmitic acid [Yabusaki, K. K., Cohen, R. E., & Ballou, C. E. (1979) J. Biol. Chem. 254, 7282-7286]. The native methylglucose lipopolysaccharide, which contains several short-chain acyl groups as well as an esterified octanoyl group, has an anomeric proton nuclear magnetic resonance spectrum similar to that of the methylglucose polysaccharide-hexadecyltrimethylammonium bromide complex. This suggests that the acylation stabilizes the polysaccharide chain in the same conformation it assumes when complexed to a long-chain lipid. Thus, acylation of the methylglucose polysaccharide could have an important role in regulating its shape and lipid-binding properties.