2024-03-29T08:45:59Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/2093652020-11-12T14:39:29Zcom_10261_34com_10261_5com_10261_86com_10261_1col_10261_413col_10261_465
Insights in the recognition of Listeria cell wall teichoic acids by a phage-derived cell wall binding module
Kalograiaki, Ioanna
Shen, Yang
Menéndez, Margarita
Prunotto, Alessio
Hernández, Noelia
Dal Peraro, Matteo
Cañada, F. Javier
Loessner, Martin
SEBBM19madrid, Madrid on 16-19th July 2019, G05-13-P7 f. -- https://congresosebbm.madrid2019.es/
Endolysins are phage-encoded peptidoglycan hydrolases able to degrade the cell-wall of bacterial hosts
from the inside and the outside. Often they are modular enzymes bearing separate recognition and
catalytic functions. Susceptibility to lytic activity depends on the selectivity of their cell wall-binding
domain (CBD) frequently a carbohydrate-recognition domain. As significant example the CBD of the
phage endolysin Ply500 (CBD500) exhibits binding patterns that correlate with structural variations in
wall teichoic acids (WTAs) of Listeria spp. their target bacteria [1]. Yet atomic scale insights in this
interaction remain challenging to obtain due to the extraordinary heterogeneity of these glycopolymers in
terms of length GlcNAc/ribitol connectivity mutable O-acetylation or further hexose substitution of the
GlcNAc unit. In this work we employed a multifaceted strategy in order to decipher the CBD500 fine
sugar specificity and WTA recognition mechanism from both the ligand and protein perspectives.
Saturation Transfer Difference (STD) NMR of CBD500-complexes with a panel of native and mutant
WTAs highlighted the importance of GlcNAc 3¿O-acetylation and the relevant contributions of hexose
substitutions in binding. ¿Blind¿ molecular docking on the X-ray structure of CBD500 allowed us to
propose the WTA binding site and the putative interacting groups further assessed by site-directed
mutagenesis. STD NMR-driven molecular dynamic simulations and isothermal titration calorimetry (ITC)
aided to model the CBD500 interaction with targeted WTA and analyze the H-bond network as well as
hydrophobic interactions established. This pioneer study unveiled the previously unknown recognition
mechanism between CBD500 and Listeria native WTA polymers thus guiding further studies aiming to
decipher the regulation of endolysin specificity.
2020-04-28T09:50:18Z
2020-04-28T09:50:18Z
2019-07-16
2020-04-28T09:50:18Z
póster de congreso
http://purl.org/coar/resource_type/c_6670
42rd Congress of the Spanish Biochemical and Molecular Biology Society (2019)
http://hdl.handle.net/10261/209365
Sí
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