Structure-function analysis of the self-recognizing Antigen 43 autotransporter protein from Escherichia coli

Per Klemm, L. Hjerrild, Morten Gjermansen, Mark Schembri

    Research output: Contribution to journalJournal articleResearchpeer-review


    Antigen 43 (Ag43) is a self-recognizing surface adhesin found in most Escherichia coli strains. Expression of Ag43 confers aggregation and fluffing of cells, promotes biofilm formation and is associated with enhanced resistance to antimicrobial agents. Ag43 is an autotransporter protein and consists of two moieties: a transporter, the beta-module, and a passenger domain, the alpha-module. Here we have employed various molecular approaches to probe structure/function aspects of Ag43. An entire family of Ag43 variants was identified. The gene encoding Ag43 (flu) was cloned from a diverse range of E. coli subtypes and found to encode variant proteins with different properties. Several novel variants were identified and characterized that were unable to promote cell-cell aggregation. By employing a combination of linker insertion mutagenesis and domain swapping between clumping and non-clumping variants, we have pinpointed the region of the protein responsible for autoaggregation to be located within the N-terminal one-third of the passenger domain. Our data suggest that ionic interactions between charged residues residing in interacting pairs of Ag43(alpha) domains may be important for the self-recognition process. Based on its similarity to other related proteins, we predict the passenger, Ag43(alpha), domain primarily to consist of an extended beta-helix structure in which numerous repeats or rungs are stacked in parallel orientation in an extended cylindrical formation. Finally, we found that in spite of their different aggregative pattern all Ag43 variants promoted biofilm formation to abiotic surfaces.
    Original languageEnglish
    JournalMolecular Microbiology
    Issue number1
    Pages (from-to)283-296
    Publication statusPublished - 2004

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