The agglutination protein AggA from Shewanella oneidensis MR-1 is a TolC-like protein and forms active channels in vitro

Type I secretion systems (TISS) are associated with the virulence of Gram-negative bacteria and the secretion of pathogenic molecular determinants. The Shewanella oneidensis MR-1 outer-membrane protein AggA is part of a TISS. Recombinant AggA expressed in Escherichia coli as inclusion bodies can be efficiently refolded in vitro, and can form active channel–tunnels as shown by proteoliposome swelling assays and electrophysiological measurements in lipid bilayers. Structure-based sequence alignments identify AggA as a TolC-like protein, and point to a conserved structural framework among such proteins despite their marginal sequence similarity. Phylogenetic analysis reveals that clustering of TolC-like proteins can be correlated with their involvement in TISS, Resistance/Nodulation/Division (RND) or Major Facilitator Superfamily (MFS) complexes. Taken together, our results establish a first set of structure–function relationships for a bacterial outer-membrane protein likely to be exclusively involved in TISS, and may contribute towards a more accurate classification of Outer-Membrane Factor (OMF) family proteins.