Unravelling the mysteries of novel two-domain lectins from
opportunistic human pathogens

a 2025

Unravelling the mysteries of novel two-domain lectins from opportunistic human pathogens

BURÁŇOVÁ, Tereza; Eva PAULENOVÁ a Michaela WIMMEROVÁ

Základní údaje

Originální název

Unravelling the mysteries of novel two-domain lectins from opportunistic human pathogens

Autoři

BURÁŇOVÁ, Tereza ; Eva PAULENOVÁ a Michaela WIMMEROVÁ

Vydání

XXI. Discussions in Structural Molecular Biology and 8th User Meeting of CIISB, 2025

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Označené pro přenos do RIV

Organizační jednotka

Přírodovědecká fakulta

ISSN

Klíčová slova česky

lektiny; dvoudoménové proteiny; rekombinantní proteiny

Klíčová slova anglicky

lectins; two-domain proteins; recombinant proteins

Změněno: 8. 4. 2026 14:54, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

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.

Návaznosti

LM2023042, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CIISB - Česká infrastruktura pro integrativní strukturní biologii

Citovat

BURÁŇOVÁ, Tereza; Eva PAULENOVÁ a Michaela WIMMEROVÁ. Unravelling the mysteries of novel two-domain lectins from opportunistic human pathogens. In XXI. Discussions in Structural Molecular Biology and 8th User Meeting of CIISB. 2025. ISSN 1211-5894.

@proceedings{2511799,
   author = {Buráňová, Tereza and Paulenová, Eva and Wimmerová, Michaela},
   booktitle = {XXI. Discussions in Structural Molecular Biology and 8th User Meeting of CIISB},
   keywords = {lectins; two-domain proteins; recombinant proteins},
   language = {eng},
   title = {Unravelling the mysteries of novel two-domain lectins from opportunistic human pathogens},
   year = {2025}
}

TY  - CONF
ID  - 2511799
AU  - Buráňová, Tereza - Paulenová, Eva - Wimmerová, Michaela
PY  - 2025
TI  - Unravelling the mysteries of novel two-domain lectins from opportunistic human pathogens
KW  - lectins
KW  - two-domain proteins
KW  - recombinant proteins
N2  - 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.
ER  -

BURÁŇOVÁ, Tereza; Eva PAULENOVÁ a Michaela WIMMEROVÁ. Unravelling the mysteries of novel two-domain lectins from opportunistic human pathogens. In \textit{XXI. Discussions in Structural Molecular Biology and 8th User Meeting of CIISB}. 2025. ISSN~1211-5894.

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