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Updated: Dec 26, 2025
Author Spotlight: Advancements in Understanding and Combatting Shigella Infections
Published on: February 9, 2024
Increasing the Affinity of an O-Antigen Polysaccharide Binding Site in Shigella flexneri Bacteriophage Sf6 Tailspike
Sonja Kunstmann1,2,3, Olof Engström4, Marko Wehle2
1Physikalische Biochemie, Universität Potsdam, Karl-Liebknecht-Str. 24-25, 14476, Potsdam, Germany.
Abstract:
Broad and unspecific use of antibiotics accelerates spread of resistances. Sensitive and robust pathogen detection is thus important for a more targeted application. Bacteriophages contain a large repertoire of pathogen-binding proteins. These tailspike proteins (TSP) often bind surface glycans and represent a promising design platform for specific pathogen sensors. We analysed bacteriophage Sf6 TSP that recognizes the O-polysaccharide of dysentery-causing Shigella flexneri to develop variants with increased sensitivity for sensor applications. Ligand polyrhamnose backbone conformations were obtained from 2D 1 H,1 H-trNOESY NMR utilizing methine-methine and methine-methyl correlations. They agreed well with conformations obtained from molecular dynamics (MD), validating the method for further predictions. In a set of mutants, MD predicted ligand flexibilities that were in good correlation with binding strength as confirmed on immobilized S. flexneri O-polysaccharide (PS) with surface plasmon resonance. In silico approaches combined with rapid screening on PS surfaces hence provide valuable strategies for TSP-based pathogen sensor design.
Insights
Bacteriophage tailspike proteins (TSP) can be engineered for sensitive pathogen detection. Molecular dynamics and surface plasmon resonance guided the development of improved Shigella flexneri sensors.
Area of Science:
- Microbiology
- Biotechnology
- Structural Biology
Background:
- Broad antibiotic use drives antimicrobial resistance, necessitating precise pathogen detection.
- Bacteriophages possess tailspike proteins (TSPs) that bind specific surface glycans, offering potential for pathogen sensor development.
- Tailspike proteins (TSPs) from bacteriophages are promising platforms for creating specific pathogen sensors.
Purpose of the Study:
- To engineer bacteriophage Sf6 TSP variants with enhanced sensitivity for detecting Shigella flexneri.
- To investigate the correlation between predicted ligand flexibility and binding strength for sensor optimization.
- To validate in silico methods for guiding TSP-based sensor design.
Main Methods:
- Utilized 2D 1H,1H-trNOESY NMR to determine polyrhamnose backbone conformations.
- Employed molecular dynamics (MD) simulations to predict ligand flexibility and conformations.
- Applied surface plasmon resonance (SPR) to confirm binding strength of TSP variants to immobilized Shigella flexneri O-polysaccharide.
Main Results:
- NMR-derived conformations closely matched MD predictions, validating the computational approach.
- MD-predicted ligand flexibilities showed a strong correlation with experimentally determined binding strengths.
- Engineered TSP variants demonstrated potential for increased sensitivity in sensor applications.
Conclusions:
- In silico approaches, including MD simulations, are valuable for predicting TSP-ligand interactions and guiding sensor design.
- Rapid screening on immobilized pathogen surfaces, like O-polysaccharide, combined with computational methods, facilitates TSP-based sensor development.
- This study provides a framework for engineering bacteriophage TSPs into sensitive and specific pathogen detection systems.
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