Analysis of the cell surface layer ultrastructure of the oral pathogen Tannerella forsythia

forsythia is decorated with a 2D crystalline surface (S-) layer, with two diVerent S-layer glycoprotein species being present. Prompted by the predicted virulence potential of the S-layer, this study focused on the analysis of the arrangement of the individual S-layer glycoproteins by a combination of microscopic, genetic, and biochemical anal-yses. The two S-layer genes are transcribed into mRNA and expressed into protein in equal amounts. The S-layer was investigated on intact bacterial cells by transmission elec-tron microscopy, by immune Xuorescence microscopy, and by atomic force microscopy. The analyses of wild-type cells revealed a distinct square S-layer lattice with an over-all lattice constant of 10.1 § 0.7 nm. In contrast, a blurred lattice with a lattice constant of 9.0 nm was found on S-layer single-mutant cells. This together with in vitro self-assembly studies using puriWed (glyco)protein species indicated their increased structural Xexibility after self-assembly and/or impaired self-assembly capability. In conjunction with TEM analyses of thin-sectioned cells, this study demonstrates the unusual case that two S-layer glyco-proteins are co-assembled into a single S-layer. Addition-ally, Xagella and pilus-like structures were observed on T. forsythia cells, which might impact the pathogenicity of this bacterium