THE PH-6 ANTIGEN OF YERSINIA-PESTIS BINDS TO BETA-1-LINKED GALACTOSYLRESIDUES IN GLYCOSPHINGOLIPIDS

The Yersinia pestis pH 6 antigen was expressed by, and purified from, Escherichia coli containing cloned psa genes. By an enzyme-linked immu nosorbence-based assay, purified pH 6 antigen bound to gangliotetraosy lceramide (GM1A), gangliotriaosylceramide (GM2A), and lactosylceramide (LC) (designations follow the nomenclature of L. Svennerholm [J, Neur ochem, 10:613-623, 1963]). Binding to GM1A, GM2A, and LC was saturable , with 50% maximal binding occurring at 498 +/- 4, 590, and 196 +/- 3 nM, respectively, Thin-layer chromatography (TLC) overlay binding conf irmed that purified pH 6 antigen bound to GM1A, GM2A, and LC and also revealed binding to hydroxylated galactosylceramide. Intact E. coli ce lls which expressed the pH 6 antigen had a specificity similar to that of purified pH 6 in the TLC overlay assay except that nonhydroxylated galactosylceramide was also bound. The binding patterns observed indi cate that the presence of beta 1-linked galactosyl residues in glycosp hingolipids is the minimum determinant required for binding of the pH 6 antigen.