|Title||Surface display of the receptor-binding domain of the F17a-G fimbrial adhesin through the autotransporter AIDA-I leads to permeability of bacterial cells.|
|Publication Type||Journal Article|
|Year of Publication||2009|
|Authors||Van Gerven, N., M. Sleutel, F. Deboeck, H. De Greve, and J-P. Hernalsteens|
|Date Published||2009 Feb|
|Keywords||Adhesins, Bacterial, Adhesins, Escherichia coli, Bacterial Outer Membrane Proteins, Cell Membrane Permeability, Escherichia coli, Escherichia coli Proteins, Microbial Viability, Protein Structure, Tertiary, Protein Transport, Recombinant Fusion Proteins, Salmonella typhimurium|
Surface exposure of antigens on bacterial cells can be critical for eliciting an effective antibody response. Therefore, we investigated the cellular localization of the fimbrial F17a-G receptor-binding domain, fused to the translocator domain of the AIDA-I autotransporter. Synthesis of the fusion protein, under the control of the L-arabinose-inducible PBAD promoter, was shown to permeabilize Escherichia coli K-12 and Salmonella enterica serovar Typhimurium cells. The presence of permeable cells interfered with several methods that are typically used to determine surface exposure of proteins, such as protease treatment and whole-cell ELISA. Double immunofluorescence microscopy, using a second antibody directed against beta-galactosidase, a bacterial protein expressed in the cytoplasm, allowed the simultaneous detection of antigen expression and permeability in individual cells.
|Alternate Journal||Microbiology (Reading, Engl.)|