Solving the mysteries of novel two-domain lectins from human pathogens

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F25%3A00141753" target="_blank" >RIV/00216224:14310/25:00141753 - isvavai.cz</a>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

Solving the mysteries of novel two-domain lectins from human pathogens

Original language description

LecB (PA-IIL) is one of two characterized 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 patients with immune deficiencies (e.g. cystic fibrosis patients)1. Several LecB homologs were described in the past, for example, lectins produced by Burkholderia cenocepacia2. Nevertheless, there are still uncharacterized LecB-like proteins in the pathogenic bacteria, some of which contain an additional domain of unknown function. Their characterization could provide insights into the mechanism of infections and lead to the development of novel approaches to disease treatment. The aim of this project is the functional and structural characterization of three potential two-domain lectins containing a LecB-like domain with 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 characterize 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 crystallization screens were performed to obtain the crystals of the separate domains. The initial hits for X-ray crystallography are currently optimized to obtain well-diffracting crystals. For the whole proteins, electron microscopy methods are planned because of the expected high dynamics of the whole system.

Czech name

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Czech description

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Type

O - Miscellaneous

CEP classification

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OECD FORD branch

10608 - Biochemistry and molecular biology

Project

<a href="/en/project/LM2023042" target="_blank" >LM2023042: Czech Infrastructure for Integrative Structural Biology</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>S - Specificky vyzkum na vysokych skolach

Publication year

2025

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů