TY  - JOUR
A1  - Kunstmann, Ruth Sonja
A1  - Engström, Olof
A1  - Wehle, Marko
A1  - Widmalm, Göran
A1  - Santer, Mark
A1  - Barbirz, Stefanie
T1  - Increasing the affinity of an O-Antigen polysaccharide binding site in Shigella flexneri bacteriophage Sf6 tailspike protein
JF  - Chemistry – A European Journal
N2  - 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 H-1,H-1-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.
KW  - carbohydrates
KW  - molecular dynamics simulations
KW  - NMR spectroscopy
KW  - protein-carbohydrate interactions
KW  - surface plasmon resonance
Y1  - 2020
U6  - https://doi.org/10.1002/chem.202000495
SN  - 0947-6539
SN  - 1521-3765
VL  - 26
IS  - 32
SP  - 7263
EP  - 7273
PB  - Wiley-VCH
CY  - Weinheim
ER  -