ArticleJournal of bacteriology2001
Display of passenger proteins on the surface of Escherichia coli K-12 by the enterohemorrhagic E. coli intimin EaeA.
Article in Journal of bacteriology, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 73 citations in OpenAlex.
- A programmableApplied and environmental microbiology · 2026Article
- Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Bacterial surface display of human lectins in Escherichia coli.Microbial biotechnology · 2024Article
- Flow cytometric evaluation of yeast-bacterial cell-cell interactions.Biotechnology and bioengineering · 2023Article
- Engineering Tropism ofACS synthetic biology · 2021Article
- Prediction of antibody structural epitopes via random peptide library screening and next generation sequencing.Journal of immunological methods · 2017Article
- Escherichia coli surface display for the selection of nanobodies.Microbial biotechnology · 2017Review
- Article
- Crystal structures of the outer membrane domain of intimin and invasin from enterohemorrhagic E. coli and enteropathogenic Y. pseudotuberculosis.Structure (London, England : 1993) · 2012Article
- Type V secretion: mechanism(s) of autotransport through the bacterial outer membrane.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2012Review
- Article
- Miniproteins as phage display-scaffolds for clinical applications.Molecules (Basel, Switzerland) · 2011Review
- Article
- In vivo enzyme immobilization by inclusion body display.Applied and environmental microbiology · 2010Article
- Strategies for successful recombinant expression of disulfide bond-dependent proteins in Escherichia coli.Microbial cell factories · 2009Article
- The autodisplay story, from discovery to biotechnical and biomedical applications.Microbiology and molecular biology reviews : MMBR · 2007Review
- Surface display of proteins by gram-negative bacterial autotransporters.Microbial cell factories · 2006Article
- Bacterial display using circularly permuted outer membrane protein OmpX yields high affinity peptide ligands.Protein science : a publication of the Protein Society · 2006Article
- The global regulator Ler is necessary for enteropathogenic Escherichia coli colonization of Caenorhabditis elegans.Infection and immunity · 2006Article
- Intimin-mediated export of passenger proteins requires maintenance of a translocation-competent conformation.Journal of bacteriology · 2005Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Intimins are members of a family of bacterial adhesins from pathogenic Escherichia coli which specifically interact with diverse eukaryotic cell surface receptors. The EaeA intimin from enterohemorrhagic E. coli O157:H7 contains an N-terminal transporter domain, which resides in the bacterial outer membrane and promotes the translocation of four C-terminally attached passenger domains across the bacterial cell envelope. We investigated whether truncated EaeA intimin lacking two carboxy-terminal domains could be used as a translocator for heterologous passenger proteins. We found that a variant of the trypsin inhibitor Ecballium elaterium trypsin inhibitor II (EETI-II), interleukin 4, and the Bence-Jones protein REI(v) were displayed on the surface of E. coli K-12 via fusion to truncated intimin. Fusion protein net accumulation in the outer membrane could be regulated over a broad range by varying the cellular amount of suppressor tRNA that is necessary for translational readthrough at an amber codon residing within the truncated eaeA gene. Intimin-mediated adhesion of the bacterial cells to eukaryotic target cells could be mimicked by surface display of a short fibrinogen receptor binding peptide containing an arginine-glycine-aspartic acid sequence motif, which promoted binding of E. coli K-12 to human platelets. Cells displaying a particular epitope sequence fused to truncated intimin could be enriched 200,000-fold by immunofluorescence staining and fluorescence-activated cell sorting in three sorting rounds. These results demonstrate that truncated intimin can be used as an anchor protein that mediates the translocation of various passenger proteins through the cytoplasmic and outer membranes of E. coli and their exposure on the cell surface. Intimin display may prove a useful tool for future protein translocation studies with interesting biological and biotechnological ramifications.
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