The Intimin periplasmic domain mediates dimerisation and binding to peptidoglycan

Intimin and Invasin are prototypical inverse (Type Ve) autotransporters and important virulence factors of enteropathogenic Escherichia coli and Yersinia spp., respectively. In addition to a C-terminal extracellular domain and a β-barrel transmembrane domain, both proteins also contain a short N- terminal periplasmic domain that, in Intimin, includes a lysin motif (LysM), which is thought to mediate binding to peptidoglycan. We show that the periplasmic domain of Intimin – but not the shorter domain of Invasin – does bind to peptidoglycan both in vitro and in vivo, but only under acidic conditions. We present the solution structure of the Intimin LysM, which has an additional, potentially functionally relevant α-helix compared to other LysMs. In contrast to previous reports, we demonstrate that the periplasmic domain of Intimin mediates dimerisation. Our data suggests that the periplasmic domain contains two dimerisation interfaces. We further show that dimerisation and peptidoglycan binding are general features of LysM-containing inverse autotransporters. The periplasmic domain could be involved in autotransport, and peptidoglycan binding may aid in resisting mechanical and chemical stress during transit through the gastrointestinal tract