The arrangement of lipopolysaccharides on the outer membrane of Yersinia enterocolitica: an electron microscopic study

Abstract

One the cell surface of smooth of Yersinia enterocolitica 75 (Ye 75 S), and electron-lucent layer was identified in thin sections after incubation with Ye 75 S-antiserum. It is adjacent to the "double track" of the outer membrane. This layer does not appear on the analogously treated rough mutant (Ye 75 R), whose lipopolysaccharide (LPS) does not contain O-specific polysaccharides. It could be shown that this layer is formed by the O-specific side chains of LPS, which are unstained by routine methods. As the side chains of LPS obviosly make it difficult to visualize the LPS-strands in the outer membrane of rough strands was investigated. On Ye 75 R, the LPS-strands could be demonstrated only after treatment of cells with polymyxin B. After short time of exposure, the LPS appeared as contigous strand-like structures (diameter roughly 60 A) on the cell surface of cells which were negatively stained or dried by the critical-point-method. After prolonging polymyxin B treatment, "blebs" or "protrusions" appeared on the surface of Ye 75 cells, which were in negatively stained preparations and in thin sections identified as partly loosened LPS. The demonstration of LPS-strands in untreated cells was possible only on a "deep rough" mutant (Ye 161-45 a R), whose LPS contains only glucosamine and KDO besides lipid A. It is suggested that the LPS form a layer of contiguous strands (mid-to-distance 62 +/- 3 A) on the surface of negatively stained cells. According to this view, on the Ye 75 S, the O-specific side chains extend from this layer into the medium.