Unravelling the mysteries of novel two-domain lectins from opportunistic human pathogens
| Autoři | |
|---|---|
| Rok publikování | 2025 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | LecB (PA-IIL) is one of two characterised lectins (saccharide-binding proteins) from the bacterium Pseudomonas aeruginosa. Both proteins (LecA and LecB) play a significant role in bacterial infection and biofilm formation in immunocompromised patients (e.g. cystic fibrosis patients) [1]. Several LecB homologs were described in the past, for example, lectins produced by Burkholderia cenocepacia [2]. Nevertheless, there are still uncharacterised LecB-like proteins in the pathogenic bacteria, some of which contain an additional domain of unknown function. Their characterisation could provide insights into the mechanism of infection and lead to the development of novel approaches for disease treatment. The aim of this project is the functional and structural characterisation of three potential two-domain lectins containing a LecB-like domain with an emphasis on their binding properties. The genes encoding these hypothetical carbohydrate-specific proteins were identified by bioinformatic analysis, cloned into expression vectors, and expressed in Escherichia coli. In addition, new gene constructs were prepared to characterise each domain separately. A variety of methods were used to investigate thermostability (nanoDSF), homogeneity (DLS, AUC) and binding properties (ITC, AUC) of the purified proteins. Several crystallisation screens were performed to obtain the crystals of the separate domains. The initial hits for X-ray crystallography are currently being optimised to obtain well diffracting crystals. For the whole proteins, electron microscopy methods are planned because of the expected high dynamics of the whole system. |
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