Increasing the Affinity of an O‐Antigen Polysaccharide Binding Site in Shigella flexneri Bacteriophage Sf6 Tailspike Protein
Open Access
- 19 March 2020
- journal article
- research article
- Published by Wiley in Chemistry – A European Journal
- Vol. 26 (32), 7263-7273
- https://doi.org/10.1002/chem.202000495
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 analyzed bacteriophage Sf6 TSP that recognizes the O‐polysaccharide of dysentery‐causing Shigella ( S. ) 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 with surface plasmon resonance. In silico approaches combined with rapid screening on PS surfaces hence provide valuable strategies for TSP‐based pathogen sensor design.Other Versions
Funding Information
- Vetenskapsrådet (2017-03703)
- Knut och Alice Wallenbergs Stiftelse
- Deutsche Forschungsgemeinschaft (BA4046/1-2)
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