Immunochemical analysis of serologically active lipids of Mycoplasma pneumoniae

Abstract

The major complement-fixing antigen of Mycoplasma pneumoniae is found in the lipid fraction of the organism. When the lipids of M. pneumoniae were fractionated by column chromatography on silicic acid, serological activity against both rabbit and human immune sera was found in two fractions, B and D. Fraction B, eluted with chloroform-methanol (9:1), was a minor component in terms of total complement-fixing activity and contained a complex of lipids which were detected in the region characteristic of phosphatidic acids by thin-layer chromatography on Silica Gel G. Fraction D, eluted with ethyl acetate-methanol (3.5:2), had approximately the same complement-fixing antigen titer as the original lipid extract and appeared as a "comet-shaped" spot between phosphatidylethanolamine and phosphatidylcholine on Silica Gel G plates charred with sulfuric acid. However, by thin-layer chromatography on Silica Gel H impregnated with sodium tetraborate, it was demonstrated that fraction D did contain multiple components, all but one of which were carbohydrate-containing lipids (giving positive reactions when sprayed with orcinol-sulfuric acid reagent). Fraction D was found to contain glycerol and phosphate in equimolar ratios but did not contain nitrogen. Two sugars were detected which migrated on paper chromatograms with glucose and galactose.